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diff --git a/static/description.css b/static/description.css
new file mode 100644
index 0000000000000000000000000000000000000000..6877e461bd3ce1dd3313b0ec0fe971b056041910
--- /dev/null
+++ b/static/description.css
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+.bfsu-page {
+ /* background-image: url('https://static.igem.wiki/teams/5004/wiki/desc-bg.png'); */
+ width: 100vw;
+ min-height: 200vh;
+ background-size: contain;
+ background-repeat: no-repeat;
+}
+
+.page-content {
+ /* position: relative; */
+ min-height: 200vh;
+ width: 100vw;
+ margin: 0 auto;
+ /* overflow: hidden; */
+}
+
+.page-content:first-child {
+ z-index: -1;
+
+}
+
+
+/* 文案æè¿°-目录相关 */
+
+.sidebar-bg-img {
+ width: 100%;
+ height: auto;
+ position: absolute;
+ z-index: 1;
+}
+
+.sidebar {
+ width: 24%;
+ padding: 30px 20px;
+ position: sticky;
+ top: 80px;
+ height: 100vh;
+ overflow-y: scroll;
+ border-right: 2px solid #ececf4;
+}
+
+.sidebar-title {
+ font-weight: bold;
+ font-size: 2.2rem;
+ margin-bottom: 30px;
+ color: #ececf4;
+}
+
+.sidebar a {
+ text-decoration: none;
+}
+
+.sidebar-a {
+ margin: 20px 0;
+
+}
+
+.sidebar-item {
+ width: 100%;
+}
+
+
+.sidebar-item-circle {
+ width: 30px;
+ height: 30px;
+ border-radius: 50%;
+ /* background-color: #eeb8c7; */
+ margin: 0 8px;
+ transition: .2s;
+ opacity: 0;
+ border: 7px solid #ececf4;
+ background-color: transparent;
+}
+
+.sidebar-item-text {
+ margin-left: 10px;
+ /* opacity: 0; */
+
+ font-size: 1.4rem;
+ color: #ececf4;
+ font-weight: bold;
+
+ transition: 0.2s;
+ font-weight: bold;
+
+ /* transform: translateX(-50px); */
+ width: calc(100% - 80px);
+}
+
+.sidebar-item:hover > .sidebar-item-circle {
+ opacity: 1;
+
+}
+
+.sidebar-item:hover > .sidebar-item-text {
+ /* transform: translateX(0); */
+ opacity: 1;
+}
+
+
+.sidebar-item-text-active {
+ transform: translateX(0);
+ opacity: 1;
+}
+
+.sidebar-item-circle-active {
+ opacity: 1;
+
+}
+
+
+/* äºŒçº§æ ‡é¢˜ */
+.sidebar-item-circle-b {
+ width: 15px;
+ height: 15px;
+ border-radius: 50%;
+ /* background-color: #eeb8c7; */
+ margin: 0 16px;
+ transition: .2s;
+ /* background-color: #62984ebd; */
+}
+
+.sidebar-item-text-b {
+ margin-left: 50px;
+
+ width: 80%;
+ font-size: 1.1rem;
+ color: #ececf4;
+
+ transition: 0.2s;
+ font-weight: bold;
+
+ /* opacity: 0;
+ transform: translateX(-50px); */
+}
+
+
+/* ä¸‰çº§æ ‡é¢˜ */
+.sidebar-item-c {
+ margin-left: 20px;
+}
+
+.sidebar-item-circle-c {
+ width: 8px;
+ height: 8px;
+ border-radius: 50%;
+ /* background-color: #eeb8c7; */
+ margin: 0 16px;
+ transition: .2s;
+ /* background-color: #62984e96; */
+}
+
+.sidebar-item-text-c {
+ margin-left: 16px;
+ width: 80%;
+ font-size: 1.3rem;
+ color: #eab5bdb7;
+ -webkit-text-stroke: 1px #fff;
+
+ transition: 0.2s;
+ font-weight: bold;
+
+ /* opacity: 0;
+ transform: translateX(-50px); */
+}
+
+
+/* æ ‡é¢˜---通用 */
+.sidebar-b,.sidebar-c {
+ margin-bottom: 8px;
+}
+
+.sidebar-item-b,.sidebar-item-c {
+ width: 100%;
+
+}
+
+
+.sidebar-item-b:hover > .sidebar-item-text-b {
+ transform: translateX(0);
+ opacity: 1;
+}
+
+.sidebar-item-c:hover > .sidebar-item-text-c {
+ transform: translateX(0);
+ opacity: 1;
+}
+
+
+/* æ ‡é¢˜ -- æ»šåŠ¨æ¡ */
+/* æ»šåŠ¨æ¡ */
+/* åˆå§‹çŠ¶æ€ä¸‹çš„滚动æ¡æ ·å¼ */
+
+/* 默认éšè—æ»šåŠ¨æ¡ */
+.sidebar::-webkit-scrollbar {
+ width: 0;
+ transition: 1s;
+}
+
+/* é¼ æ ‡æ‚¬æµ®æ—¶æ˜¾ç¤ºæ»šåŠ¨æ¡ */
+.sidebar:hover::-webkit-scrollbar {
+ width: 4px; /* 设置滚动æ¡çš„宽度 */
+ background-color: #ececf4; /* 设置滚动æ¡çš„背景颜色 */
+}
+
+.sidebar::-webkit-scrollbar-thumb {
+ background-color: #c5c5ef; /* 设置滑å—的颜色 */
+ border-radius: 2px; /* 设置滑å—的圆角 */
+}
+
+/* 滚动æ¡è½¨é“çš„æ ·å¼ */
+.sidebar::-webkit-scrollbar-track {
+ background-color: #ececf4; /* 设置轨é“的颜色 */
+}
+
+
+
+
+/* 文案æè¿°-æ£æ–‡ç›¸å…³ */
+
+
+
+.desc-content {
+ width: 60%;
+ margin: 0 auto;
+ padding: 60px 30px;
+ background-color: #dde4ec80;
+ backdrop-filter: blur(20px);
+ -webkit-backdrop-filter: blur(6px);
+ border: 0.666667px solid rgba(255, 255, 255, 0.18);
+ box-shadow: rgba(142, 142, 142, 0.19) 0px 6px 15px 0px;
+ -webkit-box-shadow: rgba(142, 142, 142, 0.19) 0px 6px 15px 0px;
+ border-radius: 12px;
+ -webkit-border-radius: 12px;
+ /* color: rgba(255, 255, 255, 0.75); */
+}
+
+.desc-content-title {
+ margin: 30px 0;
+ width: 100%;
+}
+
+.desc-content-title-circle {
+ /* margin-top: 11px;
+ height: 20px;
+ width: 20px;
+ border-radius: 50%;
+ background-color: #62984e;
+ box-shadow: 0px 0px 5px #67a84f, 0px 0px 10px #537b44; */
+}
+
+.desc-content-title-text {
+ /* margin-left: -9px; */
+ width: 64%;
+ font-size: 2rem;
+ font-weight: bold;
+}
+
+.desc-content-common-text {
+ font-size: 1.2rem !important;
+ line-height: 1.5 !important;
+ font-family: math;
+ overflow: inherit;
+ white-space: pre-wrap;
+ word-wrap: break-word;
+}
+
+
+.desc-content > img {
+ width: 80%;
+ margin: 0 auto 30px auto;
+ height: auto;
+}
+
+.desc-content ul li {
+ font-size: 1rem !important;
+ line-height: 1.5 !important;
+ font-family: math;
+ overflow: inherit;
+}
+
+.desc-content-img-desc {
+ color: #eaf3ff;
+ text-align: center;
+}
\ No newline at end of file
diff --git a/static/jquery.js b/static/jquery.js
new file mode 100644
index 0000000000000000000000000000000000000000..200b54e470a5d67925b91e87f4e769191a9f8c16
--- /dev/null
+++ b/static/jquery.js
@@ -0,0 +1,2 @@
+/*! jQuery v3.6.0 | (c) OpenJS Foundation and other contributors | jquery.org/license */
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diff --git a/static/style.css b/static/style.css
index 11599780f3d5caed7da7ff506293c0431671565a..ffb8541211ef02ad8ad5af97ae467e32fbb3c6d3 100644
--- a/static/style.css
+++ b/static/style.css
@@ -1,7 +1,125 @@
-body { padding-top: 56px; }
+body {
+ padding-top: 56px;
+}
+
+.left-aligned {
+ margin-left: auto;
+}
+
+.bg-dark {
+ background-color: #eab5bd !important;
+}
+
+.bg-hero {
+ background-color: #45b06cff;
+}
+
+/* CALLOUT */
+.bd-callout {
+ padding: 1.25rem;
+ margin-top: 1.25rem;
+ margin-bottom: 1.25rem;
+ border: 1px solid #e9ecef;
+ border-left-width: .25rem;
+ border-radius: .25rem
+}
+
+.bd-callout h4 {
+ margin-bottom: .25rem
+}
+
+.bd-callout p:last-child {
+ margin-bottom: 0
+}
+
+.bd-callout code {
+ border-radius: .25rem
+}
+
+.bd-callout+.bd-callout {
+ margin-top: -.25rem
+}
+
+.bd-callout-info {
+ border-left-color: #5bc0de
+}
+
+.bd-callout-warning {
+ border-left-color: #f0ad4e
+}
+
+.bd-callout-danger {
+ border-left-color: #d9534f
+}
+
+body {
+ overflow-x: hidden;
+ background-color: #fff0ed;
+ /* 主题色 */
+}
+
+/* FLEX 布局 */
+.flex-row {
+ display: flex;
+ flex-direction: row;
+}
+
+.flex-column {
+ display: flex;
+ flex-direction: column;
+}
+
+.align-center {
+ align-items: center;
+}
+
+.justify-between {
+ justify-content: space-between;
+}
+
+.justify-around {
+ justify-content: space-around;
+}
+
+.flex-wrap {
+ flex-wrap: wrap;
+}
+
+/* footer */
+footer {
+ position: relative;
+ z-index: 200;
+}
+
+footer a {
+ color: white;
+ font-weight: bold;
+ text-decoration: none;
+}
+
+footer a:hover {
+ color: white;
+ text-decoration: underline;
+}
+
+/* header */
+header {
+ width: 100vw;
+}
+
+header img {
+ width: 100vw;
+ height: auto;
+ margin-top: -80px;
+}
+
+body {
+ overflow-x: hidden;
+ background-color: transparent; /* 主题色 */
+}
.left-aligned { margin-left: auto; }
-.bg-dark { background-color: #343a40 !important; }
-.bg-hero { background-color: #45b06cff; }
+.bg-dark { background-color: #eab5bd !important; }
+.bg-hero { background-color: #eab5bd; }
/* CALLOUT */
.bd-callout { padding:1.25rem; margin-top:1.25rem; margin-bottom:1.25rem; border:1px solid #e9ecef; border-left-width:.25rem; border-radius:.25rem }
@@ -13,6 +131,298 @@ body { padding-top: 56px; }
.bd-callout-warning { border-left-color:#f0ad4e }
.bd-callout-danger { border-left-color:#d9534f }
+
+/* FLEX 布局 */
+.flex-row {
+ display: flex;
+ flex-direction: row;
+}
+
+.flex-column {
+ display: flex;
+ flex-direction: column;
+}
+
+.align-center {
+ align-items: center;
+}
+
+.justify-center {
+ justify-content: center;
+}
+
+.justify-between {
+ justify-content: space-between;
+}
+
+.justify-around {
+ justify-content: space-around;
+}
+
+.flex-wrap {
+ flex-wrap: wrap;
+}
+
/* footer */
footer a { color: white; font-weight: bold; text-decoration: none; }
footer a:hover { color: white; text-decoration: underline; }
+
+
+/* header */
+header {
+ width: 100vw;
+}
+
+header img {
+ width: 100vw;
+ height: auto;
+}
+
+
+
+
+
+/* home */
+
+.myContainer {
+ width: 100vw;
+ height: 1550vh;
+ display: flex;
+ justify-content: flex-start;
+ align-items: center;
+ flex-direction: column;
+}
+
+.bgImage {
+ width: 100vw;
+ position: fixed;
+ z-index: -1;
+ opacity: 0.8;
+ top: 0;
+}
+
+.imageBox {
+ width: 100vw;
+ height: 1550vh;
+ display: flex;
+ justify-content: flex-start;
+ align-items: center;
+ flex-direction: column;
+ position: absolute;
+ z-index: 10;
+}
+
+.textBox {
+ width: 100vw;
+ height: 1550vh;
+ display: flex;
+ justify-content: flex-start;
+ align-items: center;
+ flex-direction: column;
+ position: absolute;
+ z-index: 20;
+}
+
+.text1 {
+ position: absolute;
+ top: 1578px;
+ width: 500px;
+ right: 129px;
+ height: 512px;
+ font-size: 30px;
+ letter-spacing: 4px;
+}
+
+.text2 {
+ position: absolute;
+ top: 2956px;
+ width: 500px;
+ left: 246px;
+ height: 512px;
+ font-size: 30px;
+ letter-spacing: 4px;
+}
+
+.text3 {
+ position: absolute;
+ top: 4921px;
+ width: 500px;
+ right: 485px;
+ height: 512px;
+ font-size: 30px;
+ letter-spacing: 4px;
+}
+
+.text4 {
+ position: absolute;
+ top: 5942px;
+ width: 597px;
+ right: 403px;
+ height: 512px;
+ font-size: 24px;
+ letter-spacing: 4px;
+}
+
+.text5 {
+ position: absolute;
+ top: 6980px;
+ width: 597px;
+ left: 311px;
+ height: 512px;
+ font-size: 28px;
+ letter-spacing: 4px;
+}
+
+.text6 {
+ position: absolute;
+ top: 8279px;
+ width: 848px;
+ left: 339px;
+ height: 512px;
+ font-size: 28px;
+ letter-spacing: 4px;
+}
+
+.text7 {
+ position: absolute;
+ top: 9994px;
+ width: 542px;
+ right: 288px;
+ height: 512px;
+ font-size: 28px;
+ letter-spacing: 4px;
+ text-align: center;
+}
+
+.text8 {
+ position: absolute;
+ top: 10907px;
+ width: 542px;
+ right: 288px;
+ height: 512px;
+ font-size: 28px;
+ letter-spacing: 4px;
+ text-align: center;
+}
+
+/* team */
+.teamContainer {
+ width: 100vw;
+ height: 400vh;
+ display: flex;
+ justify-content: flex-start;
+ align-items: center;
+ flex-direction: column;
+}
+
+.photoBox {
+ position: absolute;
+ top: 1073px;
+ width: 1100px;
+ height: 1700px;
+ display: flex;
+ justify-content: space-around;
+ align-items: center;
+ flex-direction: row;
+ flex-wrap: wrap;
+}
+
+.photoStyle {
+ position: relative;
+ width: 300px;
+ z-index: 20;
+}
+
+.pic {
+ position: absolute;
+ z-index: 30;
+ width: 400px;
+}
+
+.nonep {
+ display: none;
+}
+
+.phoSty {
+ max-width: 200px;
+ max-height: 200px;
+ margin-top: 80px;
+}
+
+.innPic {
+ display: flex;
+ justify-content: flex-start;
+ align-items: center;
+ flex-direction: column;
+}
+
+.texPho {
+ font-size: 12px;
+ margin: 20px;
+}
+
+/* des */
+.detextBox {
+ width: 100vw;
+ z-index: 20;
+ display: flex;
+ flex-direction: row;
+ justify-content: center;
+}
+
+.deNav {
+ width: 24vw;
+ position: sticky;
+ height: 50vh;
+ top: 140px;
+ float: left;
+ padding-left: 50px;
+ bottom: 200px;
+}
+
+.deCont {
+ width: 60vw;
+ height: 400vh;
+ float: right;
+ padding-right: 60px;
+ background-color: rgba(255, 255, 255, 0.25);
+ backdrop-filter: blur(6px);
+ -webkit-backdrop-filter: blur(6px);
+ border: 0.666667px solid rgba(255, 255, 255, 0.18);
+ box-shadow: rgba(142, 142, 142, 0.19) 0px 6px 15px 0px;
+ -webkit-box-shadow: rgba(142, 142, 142, 0.19) 0px 6px 15px 0px;
+ border-radius: 12px;
+ -webkit-border-radius: 12px;
+ color: rgba(255, 255, 255, 0.75);
+}
+
+.detitle {
+ font-size: 35px;
+ font-weight: 600;
+ margin: 20px 0;
+}
+
+.nortext {
+ font-size: 20px;
+ margin: 10px 0;
+}
+
+.depicture {
+ width: 80%;
+ margin: 20px 0;
+}
+
+.navAStyle {
+ text-decoration: none;
+ color: #000;
+}
+
+.navAStyle:hover {
+ color: #000;
+}
+
+.navTitle {
+ font-size: 20px;
+ font-weight: 400;
+ color: #000;
+}
\ No newline at end of file
diff --git a/static/temp.md b/static/temp.md
new file mode 100644
index 0000000000000000000000000000000000000000..8582d4119987c78b5f05285ddadb81a3031168be
--- /dev/null
+++ b/static/temp.md
@@ -0,0 +1,151 @@
+#Experimental preparation for week 1
+Experimental plan: Prepare the medium and experimental consumables and activate the plasmid-carrying strain
+Experimental procedure:
+1. Use of ultra-clean table: open the ultra-clean table, place the centrifuge tube plate, test tube rack, Marker pen, alcohol disinfection and sterilization, and conduct UV irradiation for 30 minutes.
+2. Sterilization: Prepare Eppendorf tubes, PCR tubes, and centrifuge tubes, and wrap them with gauze. Fill 2 boxes each of large, medium, small, and gun heads, using Velcro paper to wrap the shell, and using string/rubber band to bind the gun head box. Using pre-prepared LB medium, divide 20 tubes into 50 mL x 2 bottles of LB liquid medium. The above items were sterilized using 121 ° C for 15 min (this process was about 1.5 h). An experimental accident occurred during the filling of liquid medium: the medium was spilled on the ultra-clean table, and it was cleaned up properly afterwards.
+3. Activated strains: 1 tube of E. coli DH5α/ empty vector 1 and 1 tube of DH5α/ pheP gene. One tube of E. coli DH5α/ empty vector 2 and one tube of DH5α/ carrying PAL gene. Turn on the ultra-clean table, ventilated, put the sterilized LB test tube into the table, light the alcohol lamp, burn the test tube orifice and plug, put 100 μL of preserved bacteria solution into each test tube, add 2.5 μL Amp antibiotic mother solution, burn the test tube orifice and plug again. After sealing the tube, the tube was bundled and incubated on a shaker overnight (150 rpm, 37℃).
+
+#2 weeks to extract the templates, began to construct the plasmid
+Experimental plan: Plasmid was extracted and the target fragment was amplified by PCR
+Experimental procedure:
+1. Plasmid extraction: Four kinds of plasmids DH5α/ empty vector 1 and one tube DH5α/ pheP gene were extracted according to the instruction of plasmid extraction kit. One tube of E. coli DH5α/ empty vector 2 and one tube of DH5α/ carrying PAL gene. The bacterial solution was centrifuged for 1min and then the precipitate was removed. p1 was added separately and resuspended, p2 was mixed upside down, p3 was mixed upside down, and centrifuged for 10min. 500μL BL reagent was added to the adsorption column, the waste solution was poured, the supernatant after centrifugation as described above was added, and the mixture was rinsed twice using pw rinse solution, and centrifuged for 1min each time. EB solution 50μL was added to the adsorption column membrane and centrifuged for 2min. The recovered DNA concentration was determined using Nanodrop and the DNA was stored at -20 ° C. In this step, most people successfully extracted the plasmid.
+图片
+Strains and plasmid sample white floc after drip into the P3
+
+图片
+
+The experimental work
+2. Dilute the primers
+The dry powder primer was centrifuged at 12,000 RPM for 1min, and the corresponding amount of distilled water was added to dilute the primer to the appropriate concentration
+图片
+
+Centrifuge dry powder primer manipulation
+
+3.PCR reaction:
+(1) Target genes pheP, PAL and two kinds of vector DNA fragments were amplified by high fidelity enzyme Pfu. The reaction system is as follows:
+Pfu 0.5 μL
+10xPfu buffer 5 μL
+Primer 1 1 μL
+Primer 2 1 μL
+Template (plasmid extracted above) : 1μL
+25μL dNTP Mixture
+20 μL of sterilized water
+(2) Perform the PCR amplification procedure
+At 98 ° C for 5 min
+The following 3-step reaction was performed for 30 cycles
+98℃ 10s
+55℃ 5s
+72℃ 1min/kb
+72℃ 8 min
+The plasmids, primers and products required for PCR are shown in the table below:
+
+#3 weeks agarose gel electrophoresis, plastic recycling, double enzyme, recycling
+Experimental plan: Agarose gel electrophoresis, gel recovery, double enzyme digestion, recovery
+Experimental procedure:
+1 Agarose gel electrophoresis: The above PCR products were added to a previously prepared agarose gel with 5μL DL 5000 DNA marker and subjected to electrophoresis (120V, 20 min). At the end of electrophoresis, the gel imaging system was used for observation. The target DNA gel was cut, placed in a 2 mL centrifuge tube, and labeled.
+图片
+
+Plot of gel electrophoresis results
+2. Gel recovery: The PCR product DNA gel was recovered according to the gel recovery kit instructions
+Add 500 microliters of equilibrium solution BL to the adsorption column CA2 and centrifuge for 1 min, pour out the waste liquid in the collection tube, and put the adsorption column back into the collection tube. A single band of target DNA was cut from the agarose gel into a clean centrifuge tube. Add an equal volume of solution PN to the gel block and place in a water bath at 50 ° C until the gel block is completely dissolved. The solution obtained in the previous step was added to another adsorption column CA2, placed at room temperature for 2 min, and centrifuged for 1min. Add 600 microliters of bleach solution PW to the adsorption column CA2 (please check whether absolute ethanol has been added before use), centrifuge for 1min, repeat rinsing twice, and let dry. Add an appropriate amount of elution buffer EB to the middle position of the adsorption membrane and place it at room temperature for 2 min. The DNA solution was collected by centrifugation for 2min. The recovered DNA concentration was determined using Nanodrop.
+ 图片
+
+Completed rubber recycling product concentration determination of DNA
+3. Double digestion: EcoRI and XhoI were used to double digest the extracted plasmid pET23b and the amplified target DNA fragment. 5x20 μL reaction. The 20 μL digestion system is as follows:
+1 μg of DNA
+1 μL of EcoRI
+1 μL XhoI
+4 μL 10xY buffer
+H2O up to 20μL
+The enzyme was digested at 37 ° C for 4h
+4 Agarose gel electrophoresis: The digested product was added to the agarose gel, 5μL of DL 5000 DNA marker was added, and electrophoresis was performed (120V, 20 min). At the end of electrophoresis, the gel imaging system was used for observation. The target DNA gel was cut, placed in a 2 mL centrifuge tube, and labeled. The product was extracted successfully, but the amount was small. After that, the second gum recovery was carried out.
+ 图片
+
+The researchers cut gel
+图片
+
+图片
+
+Photo of the experimenter at work
+
+#Competent cells were prepared at week 4
+Experimental plan: E. coli DH5α and Rosetta were activated to prepare competent cells
+Experimental procedure:
+1. Activated strains: including Escherichia coli DH5α and Rosetta. Turn on the ultra-clean table, ventilate, put the sterilized LB test tube into the table, light the alcohol lamp, burn the test tube mouth and test tube stopper, connect 100 μL of preserved bacteria solution into each test tube, and burn the test tube mouth and test tube stopper again. The test tube was closed, bundled, and incubated on a shaker overnight (220 rpm, 37℃).
+Experimental procedure:
+2. Preparation of competent cells:
+(1) Open the ultra-clean stage, put in 1 mL tip, 200 μL tip, 5 mL tip, 2mL and 30 mL centrifuge tubes, and 2 x 50 mL LB medium for UV irradiation. The 0.1 M cacl 2 and 15% 0.1 M cacl 2 solution were pre-cooled in a refrigerator at 4 ° C.
+(2) DH5α and Rosetta competent cells were prepared as follows: pre-activated and packed DH5α and Rosetta bacteria were precooled in a centrifuge at 4 ° C, centrifuged at 5000 g for 10 min, the supernatant was discard, and 800 microliters of 0.1 M CaCl2 solution was added to an ice bath for 40 min. After centrifugation again (4000 g, 10 min), the supernatant was discarded, and 100 microliters of 15% glycerol 0.1 M CaCl2 were added to each tube to resuspend the bacteria. Aliquots of competent cells into 2 mL centrifuge tubes of 100μL per tube were dispensed on ice. After labeling, the competent cells were stored in the refrigerator at -80℃.
+图片
+
+图片
+
+1. Pre-activated aliquots of DH5α and Rosetta bacterial samples
+2. Bacterial samples in an ice bath
+3 Add the CaCl2 solution
+4. After secondary centrifugation of the bacterial sample, white precipitate was produced
+5 Add glycerol CaCl2 mixing buffer
+
+#Week 5 Connect and transform
+Experimental plan: Prepare solid AGAR plates containing antibiotics, perform ligation reactions, and transform ligation products
+Experimental procedure:
+1. Preparation of antibiotic plates: LB solid medium (4.5 g AGAR added to 300 mL LB liquid medium) was prepared and sterilized at 121 ° C for 15 min by autoclave. After cooling to about 60℃, add 150 μL of 100mg/mL Amp antibiotic, shake evenly, and pour into the disposable Petri dish (about 10-15 plates).
+图片
+
+Pour the liquid LB medium into the disposable Petri dish
+2 Ligation reaction: The vector was ligated by double enzyme digestion of the target fragment. The ligation system is as follows (Note: This system is an empirical system, and the optimal linking vector/target fragment volume ratio can also be calculated according to the instructions) :
+T4 DNA ligase 1 μL
+2 μL of 10 x ligation buffer
+ 1 μL of Linear plasmid
+ Insert 6 μL of DNA
+ Connections were made using a PCR apparatus at 16 ° C for 1h. pET23b-insert was obtained, and the ligation product was stored at -20 ° C.
+3. Transformation of ligation products: 10 μL of the above ligation products were added to 100 μL competent cells, followed by an ice bath for 30 min, a water bath at 42 ° C for 60 s, and an ice bath for 5 min. Then, 1 mL LB liquid medium was added and incubated for 1 h for recovery (37 ° C, 150 rpm). In an ultra-clean stage, 100 μL of recovery solution was taken and the Amp antibiotic plate was coated with a sterile coating rod.
+
+#At the 6th week, the two successfully constructed plasmids were extracted and expanded
+Experimental plan: colony PCR verification, expansion of positive clones, extraction and verification of the correct plasmid and transformation
+Experimental procedure:
+1.PCR verification: Six colonies were labeled on each plate. Half of the colonies were picked and used as templates for colony PCR using Taq enzyme. The primers are TF, TR. The reaction system and procedures were carried out according to the instructions. Configure a 50 mL agarose gel, using DL 2000 DNA marker, and perform electrophoresis. The correct size of the band indicates that the plasmid was successfully constructed
+2. Expanded culture: The correct colonies were selected and added to 5 mL LB liquid medium, and 2.5 μL Amp antibiotic was added to expand the culture.
+3. Plasmid extraction: Two kinds of plasmids were extracted according to the instruction of plasmid extraction kit. The bacterial solution was centrifuged for 1min and then the precipitate was taken. p1 was added separately and resuspended, p2 was mixed upside down, p3 was mixed upside down, and centrifuged for 10min. 500μL BL reagent was added to the adsorption column, the waste solution was poured, the supernatant after centrifugation as described above was added, and the mixture was rinsed twice using pw rinse solution, and centrifuged for 1min each time. EB solution 50μL was added to the adsorption column membrane and centrifuged for 2min. The recovered DNA concentration was determined using Nanodrop and the DNA was stored at -20 ° C. In this step, most people successfully extracted the plasmid.
+4. Transformation: Verify the correct plasmid 1 transformation into expression Escherichia coli.
+
+#The ability of cells to metabolize phe was measured at the 7th week
+Experimental plan:
+Experimental procedure:
+1 Incubation, ELISA: To measure the capacity of whole cells to metabolization of phe, the PAL engineered strain was resuspended to an OD600 of 0.1 in 1 mL of Phe assay buffer (M9 0.5% glucose containing 1 mM millimoles per liter of Phe) and incubated in microtubes. The amount of Phe was then determined according to the phenylalanine ELISA kit. , the results are shown in FIG. A below.
+2. The strains were resuspended in 1 mL Phe experiment buffer (M9 0.5% glucose containing 1 mM Phe) to an OD600 of 0.1, placed in microtubes, and incubated at 37°C for 1h. TCA concentration was determined with OD290 as shown in FIG. B.
+3 Simultaneously determine the effect of pH on PAL enzyme activity, as shown in Figure C.
+
+图片
+
+#At week 8, the effect of the hypoxic promoter was tested to verify whether the hypoxic promoter could effectively activate PAL expression
+Experimental plan: To test the effect of the hypoxic promoter and PAL expression
+Experimental procedure:
+1. The tested engineering bacteria were statically cultured in a CO2 incubator (adjust the O2 concentration to 0%, 10% and 20%). After 16h of growth, the absorbance (OD600) and mRFP fluorescence intensity data were measured using a microplate reader.
+2. The tested engineered bacteria were statically cultured in a CO2 incubator (O2 concentration was adjusted to 0%, 10% and 20%). The strains were resuspended in 1 mL Phe experiment buffer (M9 0.5% glucose containing 1 mM Phe) to an OD600 of 0.1, placed in microtubes, and incubated at 37°C for 1h. TCA concentration was determined using OD290. The Phe content was measured using an Elisa kit.
+图片
+
+
+#Trans-cinnamic acid (TCA) promoter test and determination of PAL activity at week 9
+Experimental plan: Promoter testing and determination of PAL activity
+Experimental procedure:
+1 The strain was resuspended in 1 mM Phe experiment buffer to an OD600 of 0.1 and placed in a microtube, and an additional 1 mM TCA was added to the test group. The cells were incubated at 37°C for 1h. Phe content was determined according to the phenylalanine ELISA kit, and TCA content was tested using OD290.
+
+#At the 10th week, the signal amplification switch of TP901 was coupled to test the fluorescent protein expression under different TCA content
+Experimental plan: The signal amplification switch of TP901 was coupled to test the fluorescent protein expression
+Experimental procedure:
+1. The same procedure as above was used to construct the plasmid, mRFP gene was coupled to the downstream of TCA promoter pSenCA, and the fluorescent protein expression was measured under different TCA content.
+2. Fluorescence microscope verification experiment:
+(1) Sample preparation: appropriate amount (about 30 μl) of wild type Rosetta and Rosetta-engineered bacteria droplets were dropped onto the slide and covered with a cover slip.
+(2) Turn on the switch of the power control box of the high-pressure mercury lamp.
+(3) Insert the light stopper to interrupt the light path.
+(4) Preheat for 5-10 min.
+(5) Place the slide with the sample on the stage.
+(6) Select the objective lens (in order of first low power, then high power).
+(7) Rotate the turntable of the spectroscope assembly to select the spectroscope assembly for observing mRFP (red fluorescence).
+(8) Adjust the focal length through thick and thin screws.
+(9) Open the computer connected to the microscope and click on the digital imaging system software to collect digital images.
+3. Microplate reader was used to test fluorescent protein expression: 100 μL of wild-type Rosetta, recombinant strain solution was taken in an ultra-clean stage. The supernatant was added to a 96-well plate for 4-6 replicates. The detection conditions of the microplate reader were set (mRFP excitation λ: 584 nm/10 nm, emission λ: 607/10 nm), and the readings were taken. The data were recorded using Excel.
+
+
+
diff --git a/wiki/layout.html b/wiki/layout.html
index 56ea1a2d5244a2f14e165c0288a24538c692f8e7..4c91f78529b69975dcdfa191e2fa7634ae45270b 100644
--- a/wiki/layout.html
+++ b/wiki/layout.html
@@ -12,6 +12,10 @@
<!-- Custom CSS -->
<link href="{{ url_for('static', filename = 'style.css') }}" rel="stylesheet">
+ <link href="{{ url_for('static', filename = 'description.css') }}" rel="stylesheet">
+ <link href="{{ url_for('static', filename = 'aos/aos.css') }}" rel="stylesheet">
+ <script src="{{ url_for('static', filename = 'jquery.js') }}"></script>
+ <script src="{{ url_for('static', filename = 'aos/aos.js') }}"></script>
<title>{% block title %}{% endblock %} | Squirrel-CHN - iGEM 2023</title>
@@ -21,19 +25,13 @@
{% include 'menu.html' %}
<!-- Header -->
- <header class="bg-hero py-5 mb-5">
- <div class="container h-100">
- <div class="row h-100 align-items-center">
- <div class="col-lg-12">
- <h1 class="display-4 text-white mt-5 mb-2">{{ self.title() }}</h1>
- <p class="lead mb-5 text-white-50">{% block lead %}{% endblock %}</p>
- </div>
- </div>
- </div>
+ <header>
+ <img src="{% block cover_img %}{% endblock %}" />
</header>
+
<!-- Page Content -->
- <div class="container">
+ <div>
{% block page_content %}{% endblock %}
</div>
@@ -41,6 +39,66 @@
{% include 'footer.html' %}
<!-- Bootstrap Bundle with Popper -->
+ <script>
+ $(document).ready(function () {
+ // åˆå§‹åŒ–AOS并é…ç½®
+ AOS.init({
+ duration: 500, // 动画æŒç»æ—¶é—´ï¼ˆä»¥æ¯«ç§’为å•ä½ï¼‰
+ offset: 500, // å…ƒç´ è·ç¦»è§†çª—底部多少åƒç´ 时触å‘动画
+ });
+
+ });
+ </script>
+
+
+ <!-- 侧边æ 相关 -->
+ <script>
+ $(document).ready(function () {
+ $('a').on('click', function (event) {
+ if (this.hash !== '') {
+ event.preventDefault();
+ var hash = this.hash;
+ $('html, body').animate({
+ scrollTop: $(hash).offset().top - ($(window).height() - $(hash).outerHeight()) / 2
+ }, 20);
+ }
+ });
+ });
+
+ $(document).ready(function () {
+ // 获å–侧边目录ä¸çš„所有链接
+ var links = $('.sidebar a');
+
+ var links_text = $('.sidebar-item-text');
+ var links_circles = $('.sidebar-item-circle');
+ console.log(links_text)
+
+ // 检查滚动事件
+ $(window).scroll(function () {
+ // é历æ¯ä¸ªæ£æ–‡æ ‡é¢˜
+ $('.desc-content-title-text').each(function () {
+ // 获å–æ ‡é¢˜çš„ä½ç½®
+ var position = $(this).offset().top - 500;
+ // 获å–滚动æ¡çš„ä½ç½®
+ var scroll = $(window).scrollTop();
+
+ // 如果滚动æ¡åœ¨æ ‡é¢˜ä¸Šæ–¹ï¼Œæ·»åŠ 活动类
+ if (scroll >= position) {
+ var id = $(this).attr('id');
+ // 移除所有链接的活动类
+ links_text.removeClass('sidebar-item-text-active');
+ links_circles.removeClass('sidebar-item-circle-active');
+
+
+ // å°†æ´»åŠ¨ç±»æ·»åŠ åˆ°ä¸Žæ ‡é¢˜å¯¹åº”çš„é“¾æŽ¥
+ $('.sidebar a[href="#' + id + '"] .sidebar-item-text ').addClass('sidebar-item-text-active');
+ $('.sidebar a[href="#' + id + '"] .sidebar-item-circle').addClass('sidebar-item-circle-active');
+ }
+ });
+ });
+ });
+
+ </script>
<script src="{{ url_for('static', filename = 'bootstrap.bundle.min.js') }}"></script>
</body>
</html>
diff --git a/wiki/menu.html b/wiki/menu.html
index 5bddca78e2285481ee2b1dc03da938afcff4e156..1defdd86c458973f48fac7506dc4ea90417e47d0 100644
--- a/wiki/menu.html
+++ b/wiki/menu.html
@@ -1,13 +1,73 @@
-<nav class="navbar navbar-expand-lg navbar-dark bg-dark fixed-top">
- <div class="container">
+<style>
+ /* æ·»åŠ æ ·å¼ç±» */
+ .navbar {
+ display: flex;
+ width: 100%;
+ background-color: #ffffff55 !important;
+ backdrop-filter: blur(5px);
+ transition: visibility 0s, 0.6s, opacity 0.6s linear, transform 1s;
+ height: 80px;
+
+ align-items: center;
+ justify-content: center;
+
+ }
+
+ .navbar-scroll {
+ background-color: #33536c !important;
+ backdrop-filter: blur(5px);
+ transition: visibility 0s, 0.6s, opacity 0.6s linear, transform 1s;
+ box-shadow: 10px 0 5px #2128327a;
+ }
+
+ .nav-logo {
+ height: 80px;
+ /* margin-top: 40px; */
+ }
- <!---- TEAM NAME ---->
- <a class="navbar-brand" href="#">Squirrel-CHN</a>
+ .nav-item {
+ margin: 0 20px;
+ }
+</style>
+<script>
+ $(document).ready(function() {
+ // 获å–滚动æ¡ä½ç½®
+ var scrollPosition = $(window).scrollTop();
+
+ // 判æ–滚动æ¡ä½ç½®ï¼Œåˆå§‹æ—¶åº”用é€æ˜Žæ ·å¼
+ if (scrollPosition === 0) {
+ $('.navbar').removeClass('navbar-scroll');
+ } else {
+ $('.navbar').addClass('navbar-scroll');
+ }
+
+ // 监å¬æ»šåŠ¨æ¡ä½ç½®å¹¶æ·»åŠ /åˆ é™¤æ ·å¼ç±»
+ $(window).scroll(function() {
+ scrollPosition = $(this).scrollTop();
+
+ if (scrollPosition === 0) {
+ $('.navbar').removeClass('navbar-scroll');
+ } else {
+ $('.navbar').addClass('navbar-scroll');
+ }
+ });
+ });
+</script>
+
+<!-- æ·»åŠ æ»šåŠ¨æ¡ä½ç½®æ ‡è®° -->
+<div id="scroll-marker"></div>
+
+<!-- 导航æ -->
+<nav class="navbar navbar-expand-lg navbar-dark bg-dark fixed-top">
+<div class="container" style="margin: 0;padding: 0;max-width: 90vw;">
+ <a class="navbar-brand" href="#">
+ <img src="https://static.igem.wiki/teams/5013/wiki/logo.png" class="nav-logo" />
+ <span style="margin-left: 30ox;">Squirrel-CHN - iGEM 2023</span>
+ </a>
+ <button class="navbar-toggler" type="button" data-bs-toggle="collapse" data-bs-target="#navbarSupportedContent" aria-controls="navbarSupportedContent" aria-expanded="false" aria-label="Toggle navigation">
+ <span class="navbar-toggler-icon"></span>
+ </button>
- <!---- SMALL SCREEN MENU ICON ---->
- <button class="navbar-toggler" type="button" data-bs-toggle="collapse" data-bs-target="#navbarSupportedContent" aria-controls="navbarSupportedContent" aria-expanded="false" aria-label="Toggle navigation">
- <span class="navbar-toggler-icon"></span>
- </button>
<div class="collapse navbar-collapse" id="navbarSupportedContent">
<ul class="navbar-nav ml-auto left-aligned">
@@ -17,57 +77,67 @@
<a class="nav-link" href="{{ url_for('home') }}">Home</a>
</li>
- <!---- TEAM ---->
+ <!---- PROJECT ---->
<li class="nav-item dropdown">
- <a class="nav-link dropdown-toggle" href="{{ url_for('pages', page='team') }}" id="navbarDropdown" role="button" data-bs-toggle="dropdown" aria-expanded="false">
- Team
+ <a class="nav-link dropdown-toggle" href="#" id="navbarDropdown" role="button" data-bs-toggle="dropdown" aria-expanded="false">
+ Project
</a>
<ul class="dropdown-menu" aria-labelledby="navbarDropdown">
- <li><a class="dropdown-item" href="{{ url_for('pages', page='team') }}">Team</a></li>
- <li><a class="dropdown-item" href="{{ url_for('pages', page='attributions') }}">Attributions</a></li>
+ <li><a class="dropdown-item" href="{{ url_for('pages', page='description') }}">Description</a></li>
+ <li><a class="dropdown-item" href="{{ url_for('pages', page='contribution') }}">Contribution</a></li>
+
</ul>
</li>
- <!---- PROJECT ---->
<li class="nav-item dropdown">
<a class="nav-link dropdown-toggle" href="#" id="navbarDropdown" role="button" data-bs-toggle="dropdown" aria-expanded="false">
- Project
+ Wet Lab
</a>
<ul class="dropdown-menu" aria-labelledby="navbarDropdown">
- <li><a class="dropdown-item" href="{{ url_for('pages', page='contribution') }}">Contribution</a></li>
- <li><a class="dropdown-item" href="{{ url_for('pages', page='description') }}">Description</a></li>
<li><a class="dropdown-item" href="{{ url_for('pages', page='engineering') }}">Engineering</a></li>
- <li><a class="dropdown-item" href="{{ url_for('pages', page='experiments') }}">Experiments</a></li>
+ <li><a class="dropdown-item" href="{{ url_for('pages', page='proof-of-concept') }}">Proof of Concept</a></li>
+
<li><a class="dropdown-item" href="{{ url_for('pages', page='notebook') }}">Notebook</a></li>
- <li><a class="dropdown-item" href="{{ url_for('pages', page='results') }}">Results</a></li>
+ <li><a class="dropdown-item" href="{{ url_for('pages', page='experiments') }}">Experiments</a></li>
+ <li><a class="dropdown-item" href="{{ url_for('pages', page='safety') }}">Safety</a></li>
+
+
</ul>
</li>
- <!---- SAFETY ---->
- <li class="nav-item">
- <a class="nav-link" href="{{ url_for('pages', page='safety') }}">Safety</a>
- </li>
+ <li class="nav-item dropdown">
+ <a class="nav-link dropdown-toggle" href="#" id="navbarDropdown" role="button" data-bs-toggle="dropdown" aria-expanded="false">
+ Dry Lab
+ </a>
+ <ul class="dropdown-menu" aria-labelledby="navbarDropdown">
+ <li><a class="dropdown-item" href="{{ url_for('pages', page='hardware') }}">Hardware</a></li>
+ <li><a class="dropdown-item" href="{{ url_for('pages', page='model') }}">Model</a></li>
- <!---- HUMAN PRACTICES ---->
- <li class="nav-item">
- <a class="nav-link" href="{{ url_for('pages', page='human-practices') }}">Human Practices</a>
+
+ </ul>
</li>
- <!---- AWARDS ---->
<li class="nav-item dropdown">
<a class="nav-link dropdown-toggle" href="#" id="navbarDropdown" role="button" data-bs-toggle="dropdown" aria-expanded="false">
- Awards
+ Human Pratices
</a>
<ul class="dropdown-menu" aria-labelledby="navbarDropdown">
- <li><a class="dropdown-item" href="{{ url_for('pages', page='education') }}">Education</a></li>
+ <li><a class="dropdown-item" href="{{ url_for('pages', page='human-practices') }}">Human Pratices</a></li>
<li><a class="dropdown-item" href="{{ url_for('pages', page='entrepreneurship') }}">Entrepreneurship</a></li>
- <li><a class="dropdown-item" href="{{ url_for('pages', page='hardware') }}">Hardware</a></li>
- <li><a class="dropdown-item" href="{{ url_for('pages', page='inclusivity') }}">Inclusivity</a></li>
- <li><a class="dropdown-item" href="{{ url_for('pages', page='measurement') }}">Measurement</a></li>
- <li><a class="dropdown-item" href="{{ url_for('pages', page='model') }}">Model</a></li>
- <li><a class="dropdown-item" href="{{ url_for('pages', page='plant') }}">Plant</a></li>
- <li><a class="dropdown-item" href="{{ url_for('pages', page='software') }}">Software</a></li>
- <li><a class="dropdown-item" href="{{ url_for('pages', page='sustainable') }}">Sustainable</a></li>
+
+
+ </ul>
+ </li>
+
+ <!---- TEAM ---->
+ <li class="nav-item dropdown">
+ <a class="nav-link dropdown-toggle" href="{{ url_for('pages', page='team') }}" id="navbarDropdown" role="button" data-bs-toggle="dropdown" aria-expanded="false">
+ Team
+ </a>
+ <ul class="dropdown-menu" aria-labelledby="navbarDropdown">
+ <li><a class="dropdown-item" href="{{ url_for('pages', page='team') }}">Team</a></li>
+ <li><a class="dropdown-item" href="{{ url_for('pages', page='collaborations') }}">Collaborations</a></li>
+ <li><a class="dropdown-item" href="{{ url_for('pages', page='attributions') }}">Attributions</a></li>
</ul>
</li>
diff --git a/wiki/pages/attributions.html b/wiki/pages/attributions.html
index f32816b15238372c4dec07335dc21b924d44337e..4d7dd85a737f068622c094ea1463777f0a76e83d 100644
--- a/wiki/pages/attributions.html
+++ b/wiki/pages/attributions.html
@@ -1,22 +1,9 @@
{% extends "layout.html" %}
{% block title %}Attributions{% endblock %}
-{% block lead %}This page must show the attribution form of your project. This includes the work done by each of the student members on your team and any work that was done by people outside of your team, including the host labs, advisors, instructors, and individuals not on the team roster. This requirement is not about literature references - these can and should be displayed throughout your wiki.{% endblock %}
-
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/attributions.jpg{% endblock %}
{% block page_content %}
-<div class="row mt-4">
- <div class="col">
- <div class="bd-callout bd-callout-info">
- <h4>Bronze Medal Criterion #2</h4>
- <p>Describe what work your team members did and what other people did for your project.</p>
- <p>The form that bas been embded in an iframe in this page shows your team's Project Attribution form. This page must keep the form as it is.</p>
- <p>If you use a different website framework, make sure to embed the right URL for your team's form.</p>
- <hr>
- <p>Please see the <a href="https://competition.igem.org/deliverables/project-attribution">Project Attribution page</a> for more information.</p>
- </div>
- </div>
-</div>
<!--
======================================================================
@@ -26,9 +13,9 @@
WILL BE DISPLAYED ON THIS PAGE. DO NOT REMOVE IT, OTHERWISE YOU RISK OF
NOT MEETING BRONZE MEDAL CRITERION #2
-->
-<div class="row mt-4">
+<div class="row mt-4" style="width: 100vw;">
<script type="text/javascript">
- // Listen to size change and update form height
+ // Listen to size change and update form height;
window.addEventListener('message', function (e) {
const {type, data} = JSON.parse(e.data);
if (type === 'igem-attribution-form') {
diff --git a/wiki/pages/collaborations.html b/wiki/pages/collaborations.html
new file mode 100644
index 0000000000000000000000000000000000000000..060e7598fe2d224ff6fd5f65e32c258f918627ef
--- /dev/null
+++ b/wiki/pages/collaborations.html
@@ -0,0 +1,95 @@
+{% extends "layout.html" %}
+
+{% block title %}Collaborations{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/collaborations.jpg{% endblock %}
+
+{% block detail_content1 %}{% endblock %}
+{% block detail_title_a1 %}Collaboration with the BFSU-ICUnited Team on Tyrosinase Advancements{% endblock %}
+{% block detail_content2 %}
+{% endblock %}
+{% block detail_title_b1 %}Background:{% endblock %}
+{% block detail_content3 %}
+In the dynamic realm of the International Genetically Engineered Machine Competition (iGEM), collaboration stands as a cornerstone for propelling scientific endeavors. It's within this spirit of synergy that our BFSU-ICUnited team, dedicated to Bio Conditioner innovation, found common ground with the Squirrel-CHN team, pioneers in curing Phenylketonuria (PKU), centered around the critical enzyme, tyrosinase. Recognizing our shared need for this enzymatic powerhouse, we embarked on a collaborative journey aimed at maximizing resources and efficiency.
+{% endblock %}
+{% block detail_img1 %}https://static.igem.wiki/teams/5013/wiki/colla/img1.jpg{% endblock %}
+{% block detail_content4 %}
+Our collaborative efforts took root in face-to-face discussions, followed by the establishment of a WeChat group and online meetings, serving as virtual crucibles of ideas.
+
+Together, we embarked on a dual-pronged approach to enzyme synthesis, focusing on two vital components:
+
+1. Shared Laboratory Equipment and Resources:
+
+ - Mutual Chemicals: Our collaboration included the exchange of essential chemicals, including tyrosine, p-chlorophenylacetic acid (PCPA, an inhibitor of tyrosinase), TMB substrate (employed for measuring tyrosinase activity), as well as common buffers and enzyme inhibitors.
+
+ - Equipment Synergy: Beyond chemicals, we pooled our resources further, generously sharing laboratory equipment such as centrifuges and thermostatic water baths. This collaborative equipment sharing significantly facilitated our respective experimental processes.
+
+2. Ongoing Exchanges and Mutual Learning:
+
+Our collaboration extended beyond resource sharing. It became a platform for addressing challenges that both teams encountered:
+
+ - Stability Conundrums: The Bio Conditioner team grappled with the issue of tyrosinase stability under specific conditions, while the Cure PKU team faced the challenge of achieving optimal enzyme yield in mass production.
+
+
+{% endblock %}
+{% block detail_title_b2 %}In Conclusion:{% endblock %}
+{% block detail_content5 %}
+Our collaborative efforts have yielded not only substantial progress within our individual projects but also the strengthening of bonds and trust between our teams. This shared journey has not only enhanced our competitiveness in the iGEM competition but has also paved the way for greater innovations in scientific research. As we continue our partnership, we remain steadfast in our belief that our shared endeavors will lead to groundbreaking achievements and advancements in the world of genetic engineering. Together, we stand poised to make a lasting impact.
+
+{% endblock %}
+
+{% block page_content %}
+<div class="page-content flex-row">
+ <img class="bgImage" src="https://static.igem.wiki/teams/5013/wiki/desc/desc-bg.jpg">
+
+ <!-- 侧边æ -->
+ <div class="sidebar flex-column">
+
+ <!-- todo -->
+ <a class="sidebar-a" href="#a1">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a1()}}</div>
+ </div>
+ </a>
+<a class="sidebar-b" href="#b1">
+ <div class="sidebar-item-b flex-row align-center">
+ <div class="sidebar-item-circle-b">
+ </div>
+ <div class="sidebar-item-text-b">{{self.detail_title_b1()}}</div>
+ </div>
+ </a>
+<a class="sidebar-b" href="#b2">
+ <div class="sidebar-item-b flex-row align-center">
+ <div class="sidebar-item-circle-b">
+ </div>
+ <div class="sidebar-item-text-b">{{self.detail_title_b2()}}</div>
+ </div>
+ </a>
+
+ </div>
+
+ <!-- 文案æ£æ–‡ -->
+ <div class="desc-content flex-column">
+ <!-- todo -->
+ <pre class="desc-content-common-text">{{self.detail_content1()}}</pre>
+<div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a1">{{self.detail_title_a1()}}</div>
+ </div>
+<pre class="desc-content-common-text">{{self.detail_content2()}}</pre>
+<h3 id="b1">{{self.detail_title_b1()}}</h3>
+<pre class="desc-content-common-text">{{self.detail_content3()}}</pre>
+<img loading="lazy" src="{{self.detail_img1()}}" />
+<pre class="desc-content-common-text">{{self.detail_content4()}}</pre>
+<h3 id="b2">{{self.detail_title_b2()}}</h3>
+<pre class="desc-content-common-text">{{self.detail_content5()}}</pre>
+
+ </div>
+
+ </div>
+
+
+
+
+{% endblock %}
\ No newline at end of file
diff --git a/wiki/pages/contribution.html b/wiki/pages/contribution.html
index 6b8b551a304b360c129c2c58ea54822bb37a9b71..0d7fb7fa708435b0b248c3e3326669064b83a6ba 100644
--- a/wiki/pages/contribution.html
+++ b/wiki/pages/contribution.html
@@ -1,20 +1,126 @@
{% extends "layout.html" %}
{% block title %}Contribution{% endblock %}
-{% block lead %}Make a useful contribution for future iGEM teams. Use this page to document that contribution.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/contribution.jpg{% endblock %}
+
+{% block detail_content1 %}{% endblock %}
+{% block detail_title_a1 %}Introduction:{% endblock %}
+{% block detail_content2 %}In this section, we showcase our tangible contributions to the IGEM community, drawing from our experiences and solutions to the unique challenges we encountered. Our focus centered on Phenylketonuria (PKU), a condition that lacks convenient and effective treatment options. To bridge this gap, we initiated a groundbreaking microbiome therapy project for PKU, offering a promising avenue for future IGEM teams. We designed our project holistically, encompassing in-depth information on PKU, the creation of PKU-treating E. coli strains, and robust biosafety measures.
+
+{% endblock %}
+{% block detail_title_a2 %}Understanding PKU:{% endblock %}
+{% block detail_content3 %}For future IGEM teams venturing into PKU research, our project serves as a comprehensive reference. We delve into the causes, symptoms, and existing treatment options of PKU, equipping teams with a strong foundation in understanding the condition. This knowledge empowers researchers to navigate the complexities of PKU and craft innovative solutions.
+
+{% endblock %}
+{% block detail_title_a3 %}Microbiome Therapy for PKU:{% endblock %}
+{% block detail_content4 %}Our pioneering approach involves utilizing synthetic biology to engineer E. coli strains capable of treating PKU. To ensure safe and effective implementation, we provide detailed insights into the design of our microbial chassis and biosafety locks. These precautions serve as a valuable resource for teams working with genetically modified organisms, fostering a culture of biosafety within the IGEM community.
+
+{% endblock %}
+{% block detail_title_a4 %}Measuring PAL Activity:{% endblock %}
+{% block detail_content5 %}To assess the efficacy of Phenylalanine ammonia-lyase (PAL), a key component in PKU treatment, we developed three comprehensive methods for PAL activity measurement and analysis. These analytical tools offer future IGEM teams a practical means of evaluating PAL's performance in phenylalanine degradation.
+
+{% endblock %}
+{% block detail_title_a5 %}Contribution to the IGEM Community:{% endblock %}
+{% block detail_content6 %}Our project contributes tangibly to the IGEM community by presenting a novel approach to PKU treatment and delivering comprehensive resources for teams interested in this field. We aim to inspire and empower future IGEM teams to pursue innovative solutions for PKU and related challenges in synthetic biology.
+
+{% endblock %}
+{% block detail_title_a6 %}Closing Remarks:{% endblock %}
+{% block detail_content7 %}In concluding our IGEM journey, we extend our gratitude to the IGEM community for its unwavering support and the collaborative spirit that defines our shared mission. We anticipate witnessing the remarkable achievements of future IGEM teams as they build upon our contributions and continue to address pressing global issues through synthetic biology.
+
+{% endblock %}
{% block page_content %}
-<div class="row mt-4">
- <div class="col">
- <div class="bd-callout bd-callout-info">
- <h4>Bronze Medal Criterion #4</h4>
- <p>Make a useful contribution for future iGEM teams. Use this page to document that contribution.<p>
- <p>If you are making a contribution by adding information to an existing Part or creating a new Part, you must document your contribution on the Part's Main Page on the <a href="http://parts.igem.org/Main_Page">Registry</a> for your team to be eligible for this criteria. You can use this page to link to that part and include additional information about your contribution.</p>
- <hr>
- <p>Please see the <a href="https://competition.igem.org/judging/medals">2023 Medals Page</a> for more information.</p>
- </div>
+<div class="page-content flex-row">
+ <img class="bgImage" src="https://static.igem.wiki/teams/5013/wiki/desc/desc-bg.jpg">
+
+ <!-- 侧边æ -->
+ <div class="sidebar flex-column">
+
+ <!-- todo -->
+ <a class="sidebar-a" href="#a1">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a1()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a2">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a2()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a3">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a3()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a4">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a4()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a5">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a5()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a6">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a6()}}</div>
+ </div>
+ </a>
+
</div>
+
+ <!-- 文案æ£æ–‡ -->
+ <div class="desc-content flex-column">
+ <!-- todo -->
+ <pre class="desc-content-common-text">{{self.detail_content1()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a1">{{self.detail_title_a1()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content2()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a2">{{self.detail_title_a2()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content3()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a3">{{self.detail_title_a3()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content4()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a4">{{self.detail_title_a4()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content5()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a5">{{self.detail_title_a5()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content6()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a6">{{self.detail_title_a6()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content7()}}</pre>
+
+ </div>
+
</div>
+
{% endblock %}
diff --git a/wiki/pages/description.html b/wiki/pages/description.html
index c4b3aca3d694a7665169dfe9126104d542e81cb0..8ab3dc9c79b7d0d3a6cfb4aebe1e63657e732845 100644
--- a/wiki/pages/description.html
+++ b/wiki/pages/description.html
@@ -1,61 +1,354 @@
{% extends "layout.html" %}
{% block title %}Project Description{% endblock %}
-{% block lead %}Describe how and why you chose your iGEM project.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/description.jpg{% endblock %}
+
+{% block detail_content1 %}{% endblock %}
+{% block detail_title_a1 %}Introduction{% endblock %}
+{% block detail_content2 %}
+Rare diseases often receive limited attention and resources, despite the profound impact they can have on affected individuals. Many rare diseases are congenital or genetic, collectively affecting approximately 0.65% of the population. Unfortunately, most of these conditions lack curative treatments, leaving patients reliant on medications to maintain their physical and mental well-being. However, these medications are often exorbitantly priced and come with a slew of side effects, making them financially burdensome for many families, ultimately leading some to forego treatment.
+
+Through a stepwise investigation, we came across Phenylketonuria (PKU), one such rare disease. Our research revealed a group of individuals who, from birth, cannot consume common foods like milk, chicken, fish, meat, eggs, or even breast milk. For them, these seemingly ordinary foods are poison, leading to intellectual disabilities and even death upon ingestion. These patients must adhere to a lifelong, highly restricted diet, earning them the endearing term, "angels who don't partake in earthly pleasures."
+{% endblock %}
+{% block detail_img1 %}https://static.igem.wiki/teams/5013/wiki/desc/img1.jpg{% endblock %}
+{% block detail_content3 %}
+With a foundational understanding of PKU, our team developed a strong desire to further assist these "angels." Consequently, we made the resolute decision to adopt PKU as the focal point of our iGEM project. Our goal is to engineer a novel treatment approach that offers cost-effectiveness, safety, and reliability for all PKU patients.
+
+{% endblock %}
+{% block detail_title_a2 %}Phenylketonuria and Its Causes{% endblock %}
+{% block detail_content4 %}Phenylketonuria is a congenital autosomal recessive genetic disorder, classified as a common amino acid metabolic disorder, with an estimated prevalence in China of approximately 1 in 11,000. PKU remains incurable, necessitating lifelong treatment. With timely treatment and proper control, it typically does not affect natural lifespan.
+About Phenylalanine
+Phenylalanine is an essential amino acid in human metabolism, with a daily requirement for a normal child estimated at 200-500 mg. Approximately one-third is used for protein synthesis, while two-thirds are converted to tyrosine by the action of phenylalanine hydroxylase (PAH) in hepatic cells, supplying the synthesis of thyroid hormone, adrenaline, melanin, and more.
+In the hydroxylation process of phenylalanine, the involvement of the coenzyme tetrahydrobiopterin (BH4) is essential.
+Causes
+PKU results from a deficiency of phenylalanine hydroxylase (PAH) in hepatic cells in the classical mechanism, or from a lack of tetrahydrobiopterin (BH4) in the non-classical mechanism, within the phenylalanine (PA) metabolic pathway. This deficiency leads to the inability to metabolize phenylalanine (Phe), resulting in the accumulation of Phe, a deficiency in tyrosine, and the production of phenyl-lactic acid and phenylpyruvic acid, subsequently causing a series of symptoms.
+{% endblock %}
+{% block detail_img2 %}https://static.igem.wiki/teams/5013/wiki/desc/img2.png{% endblock %}
+{% block detail_content5 %}
+{% endblock %}
+{% block detail_title_a3 %}Key Symptoms{% endblock %}
+{% block detail_content6 %}Accumulation of Phenylalanine: Excessive accumulation of phenylalanine can lead to neurotoxicity, potentially causing symptoms such as hypertension, brain damage, and seizures.
+Tyrosine Deficiency: Tyrosine, originally synthesized from the metabolism of phenylalanine, plays a crucial role in the production of thyroid hormones, adrenaline, and melanin. Its deficiency can lead to intellectual disabilities, growth retardation, and light pigmentation of the skin and hair.
+Excess Phenyl-Lactic Acid and Phenylpyruvic Acid: Phenylalanine is diverted into producing higher levels of phenyl-lactic acid and phenylpyruvic acid, leading to their excretion in sweat and urine, resulting in the characteristic "musty" odor.
+{% endblock %}
+{% block detail_img3 %}https://static.igem.wiki/teams/5013/wiki/desc/img3.png{% endblock %}
+{% block detail_content7 %}
+
+
+{% endblock %}
+{% block detail_title_a4 %}Current Treatment{% endblock %}
+{% block detail_content8 %}Phenylketonuria (PKU) has three main treatment methods: a low protein diet and two medicines, KUVAN and Palynziq. PKU is caused by the inability to process phenylalanine (Phe), causing an excessive buildup of it in the patient's body. Because Phe is one of the main building blocks of protein, PKU patients need to avoid consuming proteins to decrease the accumulation hein their bodies, however, by decreasing the daily intake of protein patients risk insufficiency of other essential amino acids in proteins, raising the possibility for deficiency related conditions.
+
+There are two FDA-approved drugs for PKU: KUVAN and Palynziq, both produced by BioMarin Pharmaceutical. KUVAN is a cofactor of PAH enzyme that assists the enzyme in breaking down Phe. Palynziq injection is a enzyme substitution therapy injecting phenylalanine ammonia-lyase (PAL), an enzyme that breaks down Phe, into the patient's body. KUVAN can be administered to children over one month and adults for certain kinds of PKY while Palynziq injections can only be administered to adults with PKU.
+
+{% endblock %}
+{% block detail_img4 %}https://static.igem.wiki/teams/5013/wiki/desc/img4.png{% endblock %}
+{% block detail_content9 %}
+Both drugs have many side effects and can cause severe, even life-threatening, allergic reactions, including side effects such as headache, nasal congestion and Inflammation of the lining of the esophagus or stomach in the case of KUVAN, and rashes, itching, and swelling of face for Palynziq In addition, these drugs must be used along with a Phe-restricted diet, which would negatively affect the patient's body due to the lack of nutrition.
+{% endblock %}
+{% block detail_img5 %}https://static.igem.wiki/teams/5013/wiki/desc/img5.jpg{% endblock %}
+{% block detail_content10 %}
+{% endblock %}
+{% block detail_title_a5 %}Our Design{% endblock %}
+{% block detail_content11 %}Our design can operate in two different conditions: in vivo system, in which the bacteria work inside the body, and in vitro system, in which the bacteria works outside of the body. These two systems have the same transport system, but each have a slightly different biodegration system.
+
+Internal Transformation System
+Within our in vivo transformation system, we have meticulously divided its functions into two distinct components: the transport system and the degradation system. These two systems work in perfect harmony to fulfill the objectives of our project.
+
+{% endblock %}
+{% block detail_img6 %}https://static.igem.wiki/teams/5013/wiki/desc/img6.png{% endblock %}
+{% block detail_content12 %}<div class="desc-content-img-desc">Figure: Graphic Portrayal of In Vivo Transformation System. 1) Phe is phenylalanine. 2) PAL is phenylalanine ammonia-lyase. 3) TCA is trans-cinnamic acid. </div>
+{% endblock %}
+{% block detail_title_a6 %}Transport System:{% endblock %}
+{% block detail_content13 %}The transport system operates under the control of the T7 promoter and encompasses a critical genetic module. This module expresses the PheP transport enzyme, a specialized transporter designed to actively uptake phenylalanine (Phe) into the bacterial cells. This step is vital as it prevents Phe from being absorbed by the human body upon ingestion of our engineered bacteria.
+
+{% endblock %}
+{% block detail_title_a7 %}Degradation System:{% endblock %}
+{% block detail_content14 %}The degradation system is poised to tackle the challenge of metabolizing Phe efficiently. This system is activated once our bacteria enter the intestinal environment, characterized by low oxygen levels. The switch is triggered by these anaerobic conditions, leading to the activation of the PepT promoter and metabolic machinery.
+Within the degradation system, we've harnessed the potential of the PAL enzyme. PAL, or Phenylalanine Ammonia Lyase, plays a pivotal role by catalyzing the conversion of Phe into trans-cinnamic acid (TCA). This process marks a crucial step towards neutralizing the harmful effects of excessive Phe in the body.
+
+{% endblock %}
+{% block detail_title_a8 %}Metabolic Pathway:{% endblock %}
+{% block detail_content15 %}1ã€Intestinal Absorption: TCA, the product of phenylalanine (Phe) degradation, embarks on its journey from the intestines. Here, it enters the bloodstream through the intestinal walls, carried by blood vessels.
+2ã€Hepatic Conversion: TCA-rich blood eventually reaches the liver, where a remarkable transformation occurs. The liver enzymatically converts TCA into hippuric acid (HA), a much less harmful compound.
+3ã€Renal Excretion: With its conversion complete, HA is transported to the bladder through the bloodstream. From there, it is excreted from the body via urine, effectively eliminating excess phenylalanine in a safe and efficient manner.
+{% endblock %}
+{% block detail_img7 %}https://static.igem.wiki/teams/5013/wiki/desc/img7.png{% endblock %}
+{% block detail_content16 %}<div class="desc-content-img-desc">Figure: In vivo transformation system's steps to degrade phenylalanine (Phe). 1) Phe is the acronym phenylalanine. 2) PAL stands for phenylalanine ammonia-lyase. 3) TCA is the acronym trans-cinnamic acid. 4) HA stands for hippuric acid.</div>
+{% endblock %}
+{% block detail_title_a9 %}Why Choose Extracorporeal Transformation{% endblock %}
+{% block detail_content17 %}
+1. Psychological Resistance
+After conducting extensive surveys involving patients and their families, it became evident that there exists significant psychological resistance to the direct consumption of Escherichia coli (E. coli) bacteria. They expressed a strong preference for avoiding direct contact between the human body and these bacteria.
+Moreover, concerns were raised about the in vivo system's design, which involves degrading phenylalanine (Phe) inside the body. Many patients and their families worried whether all Phe could be fully degraded by the bacteria before the protein was absorbed by the body.
+
+2. Lack of Objectivity
+Phenylalanine (Phe) is rapidly absorbed in the body, and there's a possibility that it may be absorbed by the human body before the engineered bacteria have a chance to act. When patients self-administer treatment at home, variations in the Phe content of their food introduce uncertainty regarding the optimal dosage. If the dosage is too low, Phe may not be completely cleared, resulting in residual Phe in the body.
+Additionally, legal restrictions in some countries, including China, prohibit the intake of probiotics, making it impossible to use an in vivo treatment approach.
+
+3. Advantages
+Extracorporeal transformation, conducted entirely outside the human body, offers cost-effective options even for economically disadvantaged households. Standardized production of specialized dietary products ensures a more reliable conversion of phenylalanine.
+
+
+{% endblock %}
+{% block detail_title_a10 %}Customizing Gene Pathways for Inside and Outside the Human Body Environments{% endblock %}
+{% block detail_content18 %}To address the specificities of inside and outside the human body environments, we made certain modifications to our gene pathways. The transport system remains unchanged.
+
+{% endblock %}
+{% block detail_img8 %}https://static.igem.wiki/teams/5013/wiki/desc/img8.png{% endblock %}
+{% block detail_content19 %}Within the inside the human body environment, our degradation system employs an anoxia-inducible promoter adapted to the intestinal conditions. However, in the outside the human body setting, we utilize the TCA promoter, which responds to TCA induction to activate downstream PAL gene expression.
+
+An ensuing challenge was ensuring that our engineered bacterial strains could fully degrade phenylalanine in food. To enhance PAL gene expression further, we designed a gene circuit incorporating a positive feedback loop and irreversible switch.
+{% endblock %}
+{% block detail_img9 %}https://static.igem.wiki/teams/5013/wiki/desc/img9.png{% endblock %}
+{% block detail_content20 %}We positioned the TP901 gene downstream of the TCA promoter. TP901 is an integrase enzyme capable of recognizing specific DNA sequences and altering their order. We placed a reverse T7 promoter within TP901's recognition sites.
+
+Under normal conditions when the TCA promoter is unstimulated by TCA, TP901 remains dormant, and the T7 promoter does not activate. This design reduces the burden on bacterial cells when E. coli activation is unnecessary, such as during amplification and cultivation phases.
+
+The TCA promoter exhibits low-level basal expression, allowing it to weakly act and transcribe phenylalanine ammonia lyase (PAL) in the absence of trans-cinnamic acid (TCA). These trace amounts of PAL break down Phe into minimal TCA. The TCA generated from degradation further amplifies the TCA promoter's response, leading to transcription and translation of TP901 integrase. TP901 integrase redirects the downstream T7 promoter.
+
+Once the robust T7 promoter is activated, this gene switch remains permanently open. Consequently, more PAL is expressed, enabling the complete breakdown of Phe in food.
+
+{% endblock %}
+{% block detail_title_a11 %}implementation：{% endblock %}
+{% block detail_content21 %}
+In the In Vitro process, after introducing our product, Antiphe, into the food, E. coli absorbs phenylalanine (Phe) from the food, converting it into trans-cinnamic acid (TCA) and expelling it from the bacteria. As a result, all the Phe in the treated product becomes TCA, which can be absorbed by the human body or patients. Finally, we treat and remove the residual E. coli from the food to obtain the Phe-free final product.
+
+Target User:
+
+Our target user group consists of patients with phenylketonuria (PKU), a genetically inherited amino acid metabolism disorder where individuals cannot metabolize phenylalanine (Phe) normally, leading to severe symptoms such as brain damage. Due to this condition, this group requires special diets of low or Phe-free food. To meet the needs of PKU patients, we plan to offer a variety of Phe-free food options, including bread, cakes, cookies, and various types of food products treated with Antiphe. We believe that these products will not only help patients manage their condition but also provide a diverse choice of food and convenience that are helpful in their daily lives.
+
+Product Production:
+
+The production of Antiphe involves the use of engineered E. coli bacteria that we designed. These bacteria can absorb Phe and convert it into TCA. We will cultivate these E. coli bacteria in a controlled laboratory environment and incorporate them into the food production process. After cultivating the engineered bacteria, we will transfer them to microbial fermentation factories for larger-scale cultivation before delivering them to food producers. This way, we can mass-produce foods that are low in Phe or completely Phe-free.
+
+Product Storage and Function:
+
+Our products undergo meticulous sterilization before leaving the factory to ensure the biological safety of the products. All E. coli are eliminated, leaving only the food components resulting from the bacteria's conversion. This means that our products can be stored at room temperature for extended periods while maintaining their quality and nutritional value. Each product will be labeled with detailed storage and consumption guidelines to assist producers and consumers in correctly storing and consuming our products.
+
+Product Usage:
+
+We offer a variety of flavors and types of food to meet both the needs and preferences of PKU patients. Since our products do not contain Phe, PKU patients can consume them without concerns over worsening symptoms. However, we still recommend consulting with a doctor when consuming any new food or changing dietary habits to ensure alignment with their individual health diets and needs.
+
+Biosafety Measures:
+
+We place a high emphasis on biosafety. Our product undergoes rigorous sterilization before leaving the factory to ensure all E. coli bacteria are killed. In addition, our production process strictly adheres to all food safety and biosafety regulations to ensure that our products are safe for consumers. Furthermore, we conduct regular product quality tests to ensure quality and safety.
+
+{% endblock %}
+{% block detail_title_a12 %}Conclusion{% endblock %}
+{% block detail_content22 %}In conclusion, our iGEM project represents a significant step forward in addressing the needs of PKU patients. By utilizing the In Vitro process and engineered E. coli, we've successfully created Phe-free food options that are safe, diverse, and convenient.
+Our commitment to biosafety and product quality ensures that our offerings meet the highest standards. We envision a future where PKU patients can enjoy a broader range of foods without health concerns.
+
+
+{% endblock %}
+{% block detail_title_a13 %}References：{% endblock %}
+{% block detail_content23 %}[1]Isabella VM, Ha BN, Castillo MJ, et al. Development of a synthetic live bacterial therapeutic for the human metabolic disease phenylketonuria. Nat Biotechnol. 2018;36(9):857-864. doi:10.1038/nbt.4222
+[2]Flachbart LK, Sokolowsky S, Marienhagen J. Displaced by Deceivers: Prevention of Biosensor Cross-Talk Is Pivotal for Successful Biosensor-Based High-Throughput Screening Campaigns. ACS Synth Biol. 2019;8(8):1847-1857. doi:10.1021/acssynbio.9b00149
+[3]Chien T, Harimoto T, Kepecs B, et al. Enhancing the tropism of bacteria via genetically programmed biosensors. Nat Biomed Eng. 2022;6(1):94-104. doi:10.1038/s41551-021-00772-3
+[4]Courbet A, Endy D, Renard E, Molina F, Bonnet J. Detection of pathological biomarkers in human clinical samples via amplifying genetic switches and logic gates. Sci Transl Med. 2015;7(289):289ra83. doi:10.1126/scitranslmed.aaa3601
+[5]Binder S, Schendzielorz G, Stäbler N, et al. A high-throughput approach to identify genomic variants of bacterial metabolite producers at the single-cell level. Genome Biol. 2012;13(5):R40. Published 2012 May 28. doi:10.1186/gb-2012-13-5-r40
+[6]Pi J, Wookey PJ, Pittard AJ. Cloning and sequencing of the pheP gene, which encodes the phenylalanine-specific transport system of Escherichia coli. J Bacteriol. 1991;173(12):3622-3629. doi:10.1128/jb.173.12.3622-3629.1991
+[7]Isabella VM, Ha BN, Castillo MJ, et al. Development of a synthetic live bacterial therapeutic for the human metabolic disease phenylketonuria. Nat Biotechnol. 2018;36(9):857-864. doi:10.1038/nbt.4222
+[8]de Groot MJ, Hoeksma M, Blau N, Reijngoud DJ, van Spronsen FJ. Pathogenesis of cognitive dysfunction in phenylketonuria: review of hypotheses. Mol Genet Metab. 2010;99 Suppl 1:S86-S89. doi:10.1016/j.ymgme.2009.10.016
+[9]Romani C, Palermo L, MacDonald A, Limback E, Hall SK, Geberhiwot T. The impact of phenylalanine levels on cognitive outcomes in adults with phenylketonuria: Effects across tasks and developmental stages. Neuropsychology. 2017;31(3):242-254. doi:10.1037/neu0000336
+[10]Vockley J, Andersson HC, Antshel KM, et al. Phenylalanine hydroxylase deficiency: diagnosis and management guideline [published correction appears in Genet Med. 2014 Apr;16(4):356]. Genet Med. 2014;16(2):188-200. doi:10.1038/gim.2013.157
+[10]Longo N, Siriwardena K, Feigenbaum A, et al. Long-term developmental progression in infants and young children taking sapropterin for phenylketonuria: a two-year analysis of safety and efficacy. Genet Med. 2015;17(5):365-373. doi:10.1038/gim.2014.109
+
+{% endblock %}
{% block page_content %}
-<div class="row mt-4">
- <div class="col">
- <div class="bd-callout bd-callout-info">
- <h4>Bronze Medal Criterion #3</h4>
- <p>Describe how and why you chose your iGEM project.<p>
- <hr>
- <p>Please see the <a href="https://competition.igem.org/judging/medals">2023 Medals Page</a> for more information.</p>
- </div>
+<div class="page-content flex-row">
+ <img class="bgImage" src="https://static.igem.wiki/teams/5013/wiki/desc/desc-bg.jpg">
+
+ <!-- 侧边æ -->
+ <div class="sidebar flex-column">
+
+ <!-- todo -->
+ <a class="sidebar-a" href="#a1">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a1()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a2">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a2()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a3">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a3()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a4">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a4()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a5">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a5()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a6">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a6()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a7">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a7()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a8">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a8()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a9">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a9()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a10">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a10()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a11">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a11()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a12">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a12()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a13">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a13()}}</div>
+ </div>
+ </a>
+
</div>
-</div>
-<div class="row mt-4">
- <div class="col-lg-8">
- <h2>Description</h2>
- <hr>
- <h3>Treatment of phenylketonuria through microecological therapy</h3>
- <ul>
- <li>Strains that have been developed successfully in the lab.</li>
- <li>To the probiotic factory large-scale expansion culture, further into freeze-dried powder.</li>
- <li>Use a lyophilized probiotic powder that protects against stomach acid and is treated with a coating technique and packaged.</li>
- <li>Patients need to melt our probiotics in warm honey water or warm milk before eating.</li>
- <li>Take engineered probiotics before meals.</li>
- <li>Our engineered probiotics metabolize dietary phenylalanine (Phe) into trans-cinnamic acid in the gut, which is further metabolized by the host into hippuric acid (HA) and excreted in the urine.</li>
- <li>Reduce the abnormal accumulation of phenylalanine (Phe) in the body. To treat phenylketonuria (PKU).</li>
- </ul>
- </div>
- <div class="col-lg-4">
- <h2>Inspirations</h2>
- <hr>
- <ul>
- <li><a href="https://2022.igem.wiki/dtu-denmark/description">2022 DTU-Denmark</a></li>
- <li><a href="https://2019.igem.org/Team:ITESO_Guadalajara/Description">2019 ITESO Guadalajara</a></li>
- <li><a href="https://2020.igem.org/Team:Technion-Israel/Description">2020 Technion Israel</a></li>
- <li><a href="https://2020.igem.org/Team:Botchan_Lab_Tokyo/Description">2020 Botchan Lab Tokyo</a></li>
- <li><a href="https://2020.igem.org/Team:St_Andrews/Description">2020 St Andrews</a></li>
- <li><a href="https://2020.igem.org/Team:MIT/Description">2020 MIT</a></li>
- </ul>
- </div>
-</div>
+ <!-- 文案æ£æ–‡ -->
+ <div class="desc-content flex-column">
+ <!-- todo -->
+ <pre class="desc-content-common-text">{{self.detail_content1()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a1">{{self.detail_title_a1()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content2()}}</pre>
+ <img loading="lazy" src="{{self.detail_img1()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content3()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a2">{{self.detail_title_a2()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content4()}}</pre>
+ <img loading="lazy" src="{{self.detail_img2()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content5()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a3">{{self.detail_title_a3()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content6()}}</pre>
+ <img loading="lazy" src="{{self.detail_img3()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content7()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a4">{{self.detail_title_a4()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content8()}}</pre>
+ <img loading="lazy" src="{{self.detail_img4()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content9()}}</pre>
+ <img loading="lazy" src="{{self.detail_img5()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content10()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a5">{{self.detail_title_a5()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content11()}}</pre>
+ <img loading="lazy" src="{{self.detail_img6()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content12()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a6">{{self.detail_title_a6()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content13()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a7">{{self.detail_title_a7()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content14()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a8">{{self.detail_title_a8()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content15()}}</pre>
+ <img loading="lazy" src="{{self.detail_img7()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content16()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a9">{{self.detail_title_a9()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content17()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a10">{{self.detail_title_a10()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content18()}}</pre>
+ <img loading="lazy" src="{{self.detail_img8()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content19()}}</pre>
+ <img loading="lazy" src="{{self.detail_img9()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content20()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a11">{{self.detail_title_a11()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content21()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a12">{{self.detail_title_a12()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content22()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a13">{{self.detail_title_a13()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content23()}}</pre>
-<div class="row mt-4">
- <div class="col-lg-8">
- <h2>Some advice</h2>
- <hr>
- <p>We encourage you to put up a lot of information and content on your wiki, but we also encourage you to include summaries as much as possible. If you think of the sections in your project description as the sections in a publication, you should try to be concise, accurate, and unambiguous in your achievements. Your Project Description should include more information than your project abstract.</p>
- </div>
- <div class="col-lg-4">
- <h2>References</h2>
- <hr>
- <p>iGEM teams are encouraged to record references you use during the course of your research. They should be posted somewhere on your wiki so that judges and other visitors can see how you thought about your project and what works inspired you.</p>
</div>
+
</div>
{% endblock %}
diff --git a/wiki/pages/education.html b/wiki/pages/education.html
index 80ff33dd0d5794c887ba26bb12a60734e8f81513..fc5a5bfc9f982ff45c5da15d7f56401b45af3217 100644
--- a/wiki/pages/education.html
+++ b/wiki/pages/education.html
@@ -1,7 +1,7 @@
{% extends "layout.html" %}
{% block title %}Education{% endblock %}
-{% block lead %}Innovative educational tools and outreach activities have the ability to establish a two-way dialogue with new communities by discussing public values and the science behind synthetic biology.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/education.jpg{% endblock %}
{% block page_content %}
diff --git a/wiki/pages/engineering.html b/wiki/pages/engineering.html
index a3f04a226ea88ee35129f42b6f08b174a5ddecbf..733f9b8421aa556f53c3a9e0e7d066cc2f77b7be 100644
--- a/wiki/pages/engineering.html
+++ b/wiki/pages/engineering.html
@@ -1,20 +1,305 @@
{% extends "layout.html" %}
{% block title %}Engineering Success{% endblock %}
-{% block lead %}Demonstrate engineering success in a part of your project by going through at least one iteration of the engineering design cycle.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/engineering.jpg{% endblock %}
+
+{% block detail_content1 %}{% endblock %}
+{% block detail_title_a1 %}Overview{% endblock %}
+{% block detail_content2 %}Synthetic biology is an interdisciplinary field that combines biology and engineering, aiming to design and construct new biological systems and functions. It goes beyond studying the complexity of organisms in nature and seeks to optimize or reconstruct biological systems through artificial design, exploiting the programmability of life. This programmability of biological systems provides opportunities to address global challenges such as climate change, disease treatment, and food security.
+
+In our project, we utilize a variety of components, including those previously commonly used or rarely employed, and test them multiple times to ensure their effectiveness. We have undergone numerous iterations to achieve microbial targeted therapy for diseases. The following provides detailed descriptions of the test results for each component, demonstrating the practical viability of our product.
+
+{% endblock %}
+{% block detail_title_a2 %}Cycle 1 : Initial E. coli engineering aimed to absorb Phe faced a big challenge.{% endblock %}
+{% block detail_content3 %}{% endblock %}
+{% block detail_title_b1 %}1. Design{% endblock %}
+{% block detail_content4 %}We aim to reduce the absorption of phenylalanine in the human body by using E. coli to absorb the phenylalanine ingested by patients. We have generated a transport system by introducing a gene expressing the PheP transporter. Simultaneously, we express PAL, an important enzyme that catalyzes the deamination of phenylalanine (Phe) and removes the amino group that cannot be processed by PKU patients.
+
+{% endblock %}
+{% block detail_img1 %}https://static.igem.wiki/teams/5013/wiki/engi/img1.png{% endblock %}
+{% block detail_content5 %}<div class="desc-content-img-desc">Figure 1 Schematic representation of the overall principle of Phep and PAL.</div>
+{% endblock %}
+{% block detail_title_b2 %}2. Build{% endblock %}
+{% block detail_content6 %}In this section, we constructed engineered strains expressing both PheP and PAL to produce transgenic E. coli capable of absorbing phenylalanine and degrading Phe. We used a constitutive promoter to express the gene of PheP and PAL.
+
+{% endblock %}
+{% block detail_title_b3 %}3. Test{% endblock %}
+{% block detail_content7 %}After expressing the engineered strains,measured Phe levels using a phenylalanine ELISA kit, while TCA concentration was determined by measuring OD290 with an microplate reader. The results, as shown in Figure 2A and 2B, demonstrate that after PAL expression, Phe levels decreased significantly, while TCA levels increased, indicating that PAL can effectively degrade Phe into TCA.
+{% endblock %}
+{% block detail_img2 %}https://static.igem.wiki/teams/5013/wiki/engi/img2.png{% endblock %}
+{% block detail_content8 %}<div class="desc-content-img-desc">Figure 2 Expression diagram of Phep and PAL.</div>
+
+{% endblock %}
+{% block detail_title_b4 %}4. Learn{% endblock %}
+{% block detail_content9 %}Through this test, it is known that there is an excessive amount of phenylalanine in E. coli cells, approaching a saturated state, which prevents the cells from further absorbing phenylalanine from the intestine. Therefore, it is considered to add a metabolic system led by phenylalanine deaminase.
+
+
+{% endblock %}
+{% block detail_title_a3 %}Cycle 2 : Utilize PAL to metabolize Phe.{% endblock %}
+{% block detail_content10 %}{% endblock %}
+{% block detail_title_b5 %}1. Design{% endblock %}
+{% block detail_content11 %}Although our first version of the plan partially transferred phenylpyruvic acid into the bacterial cells, the capacity of bacteria is limited. When the production of Phe exceeds the tolerance of the bacteria, it still leads to intracellular Phe saturation and accumulation in the patient's body. Therefore, our second version of the plan replaced the promoter with the anaerobic promoter pPepT. When the engineered bacteria enter the patient's gut, the anaerobic environment activates the inducible anaerobic promoter, and the PAL gene starts to express. The engineered bacteria secrete PAL, which metabolizes Phe in the patient's body into trans-cinnamic acid (TCA). TCA is then transported through the intestinal capillaries into the liver via the bloodstream, where it further metabolizes into hippuric acid (HA), ultimately being excreted in the urine.
+{% endblock %}
+{% block detail_img3 %}https://static.igem.wiki/teams/5013/wiki/engi/img3.png{% endblock %}
+{% block detail_content12 %}<div class="desc-content-img-desc">Figure3 Hypoxia-induced promoter pPepT drives PAL expression effect.</div>{% endblock %}
+{% block detail_title_b6 %}2. Build{% endblock %}
+{% block detail_content13 %}In pSB1A3 vector, we placed PAL and Phep genes downstream of the hypoxia-induced promoter pPepT. After enzymatic cleavage, ligation, and transformation, they will be successfully transferred into Escherichia coli bacteria, allowing the target genes to be expressed and producing engineered bacteria capable of transporting phenylpyruvic acid and metabolizing phenylalanine into trans-cinnamic acid.
+{% endblock %}
+{% block detail_title_b7 %}3. Test{% endblock %}
+{% block detail_content14 %}We tested the expression of the hypoxia-induced promoter, as shown in Figure 4A, and found that it was expressed normally under anaerobic conditions. Similarly, we measured the levels of Phe and TCA, as shown in Figures 4B and 4C. We observed higher TCA production under lower oxygen concentrations, indicating that PAL can play a significant role under anaerobic conditions. Furthermore, we validated the effect of environmental pH on PAL metabolic capacity, as shown in Figure 4D. PAL activity increased with increasing pH within the range of pH 5 to 8, with pH 8 being the optimal pH.
+
+{% endblock %}
+{% block detail_img4 %}https://static.igem.wiki/teams/5013/wiki/engi/img4.png{% endblock %}
+{% block detail_content15 %}<div class="desc-content-img-desc">Figure 4 Expression data graph of the anaerobic promoter.</div>{% endblock %}
+{% block detail_title_b8 %}4. Learn{% endblock %}
+{% block detail_content16 %}By comparison, we can conclude that the transformation ability of phe in bacteria containing Phep alone is weaker than that in bacteria containing both PheP and PAL. Therefore, it can be demonstrated that the inclusion of the metabolic system aligns with the expectations of the experiment.
+
+{% endblock %}
+{% block detail_title_a4 %}Cycle 3 : Degradation of Phe in food in vitro.{% endblock %}
+{% block detail_content17 %}{% endblock %}
+{% block detail_title_b9 %}1. Design {% endblock %}
+{% block detail_content18 %}Considering that it is not feasible to directly administer Escherichia coli and that in vivo medication does not comply with local regulations on food and drug safety in China, we plan to design an ex vivo transformation system. This system includes the T7 promoter, RBS ribosome-binding site, PHEP transport protein, terminator, TCA promoter, TP901 integrase, and PAL phenylalanine ammonia-lyase. We retained the design of the transport protein gene and the overall structure of the PAL enzyme from the in vivo transformation system but made modifications to the promoter region of the PAL gene sequence. We modified the deoxy promoter to the T7 promoter and reversed its direction, adding the TP901 integrase sequence and TCA promoter before it. Prior to the entire translation process, a small amount of PAL enzyme had already been translated, converting some Phe to trans-cinnamic acid. Trans-cinnamic acid promotes the action of the TCA promoter, initiating the translation of the TP901 integrase. The TP901 integrase recognizes the attB/attP site，and enables the T7 promoter to reverse and face the PAL gene sequence, initiating the translation of PAL. The strong T7 promoter rapidly produces a large amount of PAL enzyme without inhibiting bacterial growth，the schematic diagram is shown in Figure 5.
+{% endblock %}
+{% block detail_img5 %}https://static.igem.wiki/teams/5013/wiki/engi/img5.png{% endblock %}
+{% block detail_content19 %}<div class="desc-content-img-desc">Figure 5 Schematic representation of the overall principle of PAL gene expression.</div>{% endblock %}
+{% block detail_title_b10 %}2. Build{% endblock %}
+{% block detail_content20 %}We designed a trans-cinnamic acid (TCA) promoter and TP901 amplification signal switch system to efficiently express PAL in an in vitro transformation system.
+
+{% endblock %}
+{% block detail_title_b11 %}3. Test{% endblock %}
+{% block detail_content21 %} The schematic diagram of the trans-cinnamic acid (TCA) promoter is shown in Figure 6A, and the schematic diagram of the trans-cinnamic acid (TCA) promoter and TP901 amplification signal switch system is shown in Figure 6B. Firstly, we constructed mRFP downstream of the pTCA promoter and detected its expression, as shown in Figure 6C. After adding 1 mM TCA, there was a significant increase in mRFP fluorescence intensity. Secondly, we measured the content of Phe and TCA in the engineered strain using the same method, as shown in Figure 6D and E. After adding 1mM TCA, the Phe content decreased from around 1 mM to around 0 mM, while the TCA content increased from 0 mM to around 2 mM. Therefore, pTCA has a good activation ability for PAL expression.
+ To verify if the TP901 amplification signal switch system can promote gene expression downstream of pSenCA (TCA responsive promoter), we coupled mRFP downstream of pSenCA and added the TP901 amplification signal switch system. We tested the expression of fluorescent protein at different TCA concentrations. From Figure 6F, it can be observed that in an environment with 1mM TCA, the strain with the TP901 signal amplification switch system added downstream of pTCA had a fluorescence intensity of around 40 A.U, which was significantly higher compared to the control group. This indicates that TP901 has a significant effect in promoting downstream gene expression.
+{% endblock %}
+{% block detail_img6 %}https://static.igem.wiki/teams/5013/wiki/engi/img6.png{% endblock %}
+{% block detail_content22 %}<div class="desc-content-img-desc">Figure 6 Utilizing the trans-cinnamic acid (TCA) biosensor pSenCA and signal amplification switch for efficient gene expression.</div>{% endblock %}
+{% block detail_title_b12 %}4. Learn{% endblock %}
+{% block detail_content23 %}By analyzing the results, this design can basically meet the demand, but there is still the problem of slow degradation time. Therefore, we plan to use this technology in the factory production stage to produce edible products without phenylalanine.
+{% endblock %}
+{% block detail_title_a5 %}Conclusion{% endblock %}
+{% block detail_content24 %}Through three iterations of microbes, we have obtained an engineered microorganism that can rapidly degrade Phe and complies with regulations. These iterations are based on our exploration and research on the market, patients, and regional regulations.
+{% endblock %}
+{% block detail_title_a6 %}References{% endblock %}
+{% block detail_content25 %}1. Chien, Tiffany, et al. "Enhancing the tropism of bacteria via genetically programmed biosensors." Nature biomedical engineering 6.1 (2022): 94-104.
+2. Courbet, Alexis, et al. "Detection of pathological biomarkers in human clinical samples via amplifying genetic switches and logic gates." Science translational medicine 7.289 (2015): 289ra83-289ra83.
+3. Flachbart, Lion Konstantin, Sascha Sokolowsky, and Jan Marienhagen. "Displaced by deceivers: prevention of biosensor cross-talk is pivotal for successful biosensor-based high-throughput screening campaigns." ACS synthetic biology 8.8 (2019): 1847-1857.
+4. Binder, Stephan, et al. "A high-throughput approach to identify genomic variants of bacterial metabolite producers at the single-cell level." Genome biology 13.5 (2012): 1-12.
+5. Pi, Jing, PETER J. Wookey, and A. J. Pittard. "Cloning and sequencing of the pheP gene, which encodes the phenylalanine-specific transport system of Escherichia coli." Journal of bacteriology 173.12 (1991): 3622-3629.
+6. Isabella, Vincent M., et al. "Development of a synthetic live bacterial therapeutic for the human metabolic disease phenylketonuria." Nature biotechnology 36.9 (2018): 857-864.
+
+{% endblock %}
{% block page_content %}
-<div class="row mt-4">
- <div class="col">
- <div class="bd-callout bd-callout-info">
- <h4>Silver Medal Criterion #1</h4>
- <p>Demonstrate engineering success in a part of your project by going through at least one iteration of the engineering design cycle. This achievement should be distinct from your Contribution for Bronze.<p>
- <p>If you plan to show engineering success by creating a new Part that has been shown to work as expected, you must document your contribution on the Part's Main Page on the <a href="http://parts.igem.org/Main_Page">Registry</a> for your team to be eligible for this criteria.</p>
- <hr>
- <p>Please see the <a href="https://competition.igem.org/judging/medals">2023 Medals Page</a> for more information.</p>
- </div>
+<div class="page-content flex-row">
+ <img class="bgImage" src="https://static.igem.wiki/teams/5013/wiki/desc/desc-bg.jpg">
+
+ <!-- 侧边æ -->
+ <div class="sidebar flex-column">
+
+ <!-- todo -->
+ <a class="sidebar-a" href="#a1">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a1()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a2">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a2()}}</div>
+ </div>
+ </a>
+<a class="sidebar-b" href="#b1">
+ <div class="sidebar-item-b flex-row align-center">
+ <div class="sidebar-item-circle-b">
+ </div>
+ <div class="sidebar-item-text-b">{{self.detail_title_b1()}}</div>
+ </div>
+ </a>
+<a class="sidebar-b" href="#b2">
+ <div class="sidebar-item-b flex-row align-center">
+ <div class="sidebar-item-circle-b">
+ </div>
+ <div class="sidebar-item-text-b">{{self.detail_title_b2()}}</div>
+ </div>
+ </a>
+<a class="sidebar-b" href="#b3">
+ <div class="sidebar-item-b flex-row align-center">
+ <div class="sidebar-item-circle-b">
+ </div>
+ <div class="sidebar-item-text-b">{{self.detail_title_b3()}}</div>
+ </div>
+ </a>
+<a class="sidebar-b" href="#b4">
+ <div class="sidebar-item-b flex-row align-center">
+ <div class="sidebar-item-circle-b">
+ </div>
+ <div class="sidebar-item-text-b">{{self.detail_title_b4()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a3">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a3()}}</div>
+ </div>
+ </a>
+<a class="sidebar-b" href="#b5">
+ <div class="sidebar-item-b flex-row align-center">
+ <div class="sidebar-item-circle-b">
+ </div>
+ <div class="sidebar-item-text-b">{{self.detail_title_b5()}}</div>
+ </div>
+ </a>
+<a class="sidebar-b" href="#b6">
+ <div class="sidebar-item-b flex-row align-center">
+ <div class="sidebar-item-circle-b">
+ </div>
+ <div class="sidebar-item-text-b">{{self.detail_title_b6()}}</div>
+ </div>
+ </a>
+<a class="sidebar-b" href="#b7">
+ <div class="sidebar-item-b flex-row align-center">
+ <div class="sidebar-item-circle-b">
+ </div>
+ <div class="sidebar-item-text-b">{{self.detail_title_b7()}}</div>
+ </div>
+ </a>
+<a class="sidebar-b" href="#b8">
+ <div class="sidebar-item-b flex-row align-center">
+ <div class="sidebar-item-circle-b">
+ </div>
+ <div class="sidebar-item-text-b">{{self.detail_title_b8()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a4">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a4()}}</div>
+ </div>
+ </a>
+<a class="sidebar-b" href="#b9">
+ <div class="sidebar-item-b flex-row align-center">
+ <div class="sidebar-item-circle-b">
+ </div>
+ <div class="sidebar-item-text-b">{{self.detail_title_b9()}}</div>
+ </div>
+ </a>
+<a class="sidebar-b" href="#b10">
+ <div class="sidebar-item-b flex-row align-center">
+ <div class="sidebar-item-circle-b">
+ </div>
+ <div class="sidebar-item-text-b">{{self.detail_title_b10()}}</div>
+ </div>
+ </a>
+<a class="sidebar-b" href="#b11">
+ <div class="sidebar-item-b flex-row align-center">
+ <div class="sidebar-item-circle-b">
+ </div>
+ <div class="sidebar-item-text-b">{{self.detail_title_b11()}}</div>
+ </div>
+ </a>
+<a class="sidebar-b" href="#b12">
+ <div class="sidebar-item-b flex-row align-center">
+ <div class="sidebar-item-circle-b">
+ </div>
+ <div class="sidebar-item-text-b">{{self.detail_title_b12()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a5">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a5()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a6">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a6()}}</div>
+ </div>
+ </a>
+
+ </div>
+
+ <!-- 文案æ£æ–‡ -->
+ <div class="desc-content flex-column">
+ <!-- todo -->
+ <pre class="desc-content-common-text">{{self.detail_content1()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a1">{{self.detail_title_a1()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content2()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a2">{{self.detail_title_a2()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content3()}}</pre>
+ <h3 id="b1">{{self.detail_title_b1()}}</h3>
+ <pre class="desc-content-common-text">{{self.detail_content4()}}</pre>
+ <img loading="lazy" src="{{self.detail_img1()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content5()}}</pre>
+ <h3 id="b2">{{self.detail_title_b2()}}</h3>
+ <pre class="desc-content-common-text">{{self.detail_content6()}}</pre>
+ <h3 id="b3">{{self.detail_title_b3()}}</h3>
+ <pre class="desc-content-common-text">{{self.detail_content7()}}</pre>
+ <img loading="lazy" src="{{self.detail_img2()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content8()}}</pre>
+ <h3 id="b4">{{self.detail_title_b4()}}</h3>
+ <pre class="desc-content-common-text">{{self.detail_content9()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a3">{{self.detail_title_a3()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content10()}}</pre>
+ <h3 id="b5">{{self.detail_title_b5()}}</h3>
+ <pre class="desc-content-common-text">{{self.detail_content11()}}</pre>
+ <img loading="lazy" src="{{self.detail_img3()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content12()}}</pre>
+ <h3 id="b6">{{self.detail_title_b6()}}</h3>
+ <pre class="desc-content-common-text">{{self.detail_content13()}}</pre>
+ <h3 id="b7">{{self.detail_title_b7()}}</h3>
+ <pre class="desc-content-common-text">{{self.detail_content14()}}</pre>
+ <img loading="lazy" src="{{self.detail_img4()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content15()}}</pre>
+ <h3 id="b8">{{self.detail_title_b8()}}</h3>
+ <pre class="desc-content-common-text">{{self.detail_content16()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a4">{{self.detail_title_a4()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content17()}}</pre>
+ <h3 id="b9">{{self.detail_title_b9()}}</h3>
+ <pre class="desc-content-common-text">{{self.detail_content18()}}</pre>
+ <img loading="lazy" src="{{self.detail_img5()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content19()}}</pre>
+ <h3 id="b10">{{self.detail_title_b10()}}</h3>
+ <pre class="desc-content-common-text">{{self.detail_content20()}}</pre>
+ <h3 id="b11">{{self.detail_title_b11()}}</h3>
+ <pre class="desc-content-common-text">{{self.detail_content21()}}</pre>
+ <img loading="lazy" src="{{self.detail_img6()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content22()}}</pre>
+ <h3 id="b12">{{self.detail_title_b12()}}</h3>
+ <pre class="desc-content-common-text">{{self.detail_content23()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a5">{{self.detail_title_a5()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content24()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a6">{{self.detail_title_a6()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content25()}}</pre>
+
</div>
+
</div>
+
{% endblock %}
diff --git a/wiki/pages/entrepreneurship.html b/wiki/pages/entrepreneurship.html
index 8d7e5d835847def1a89bbd674548f624587e3086..7cfe0c2f28531c999a7b8e766aef8fc60cb8f666 100644
--- a/wiki/pages/entrepreneurship.html
+++ b/wiki/pages/entrepreneurship.html
@@ -1,7 +1,7 @@
{% extends "layout.html" %}
{% block title %}Entrepreneurship{% endblock %}
-{% block lead %}The entrepreneurship prize recognizes exceptional effort to build a business case and commercialize an iGEM project.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/entrepreneurship.jpg{% endblock %}
{% block page_content %}
diff --git a/wiki/pages/experiments.html b/wiki/pages/experiments.html
index cb46c0f1e5409ea306f25ebf6779b2f057e9ea40..091a6dc2d19b74189b0c44d2bd40c5de993c87d2 100644
--- a/wiki/pages/experiments.html
+++ b/wiki/pages/experiments.html
@@ -1,29 +1,23 @@
{% extends "layout.html" %}
{% block title %}Experiments{% endblock %}
-{% block lead %}Describe the research, experiments, and protocols you used in your iGEM project.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/experiments.jpg{% endblock %}
{% block page_content %}
-<div class="row mt-4">
- <div class="col-lg-8">
- <h2>What should this page contain?</h2>
- <hr>
- <p>Describe the research, experiments, and protocols you used in your iGEM project. These should be detailed enough for another team to repeat your experiments.</p>
- <p>If you made Parts this year, please remember to put all information, characterization, and measurement data on the Part's Main Page on the <a href="http://parts.igem.org/Main_Page">Registry</a>.</p>
- </div>
- <div class="col-lg-4">
- <h2>Inspirations</h2>
- <hr>
- <ul>
- <li><a href="https://2019.igem.org/Team:Nantes/Experiments">2019 Nantes</a></li>
- <li><a href="https://2019.igem.org/Team:TU_Eindhoven/Experiments">2019 TU Eindhoven</a></li>
- <li><a href="https://2019.igem.org/Team:Mingdao/Demonstrate">2019 Mingdao</a></li>
- <li><a href="https://2020.igem.org/Team:Amsterdam/Experiments">2020 Amsterdam</a></li>
- <li><a href="https://2020.igem.org/Team:NCTU_Formosa/Experiments">2020 NCTU Formosa</a></li>
- <li><a href="https://2020.igem.org/Team:USAFA/Experiments">2020 USAFA</a></li>
- </ul>
+
+<div style="min-height: 100vh;" class="page-content flex-row">
+ <img class="bgImage" src="https://static.igem.wiki/teams/5013/wiki/desc/desc-bg.jpg">
+
+ <!-- 文案æ£æ–‡ -->
+ <div style="min-height: 100vh;" class="desc-content flex-column">
+ <!-- todo -->
+<iframe src="https://static.igem.wiki/teams/5013/wiki/experi/2-experimental-protocol.pdf" width="100%" height="700px" type="application/pdf" allowfullscreen></iframe>
+
+
</div>
+
</div>
+
{% endblock %}
diff --git a/wiki/pages/hardware.html b/wiki/pages/hardware.html
index dbf97de92a6286a53b7c6d03e575efe5626c3402..5a475b8918add77ff03b3dff228eac25d8c5c400 100644
--- a/wiki/pages/hardware.html
+++ b/wiki/pages/hardware.html
@@ -1,7 +1,7 @@
{% extends "layout.html" %}
{% block title %}Hardware{% endblock %}
-{% block lead %}Hardware in iGEM should make synthetic biology based on standard parts easier, faster, better, or more accessible to our community.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/hardware.jpg{% endblock %}
{% block page_content %}
diff --git a/wiki/pages/home.html b/wiki/pages/home.html
index 99c8a76e75089804c36699ac4db4c4e0674e08b3..e27dc72acbd001a2503c73716e91b3d401dd672d 100644
--- a/wiki/pages/home.html
+++ b/wiki/pages/home.html
@@ -1,7 +1,7 @@
{% extends "layout.html" %}
{% block title %}Home{% endblock %}
-{% block lead %}<b>Welcome to iGEM 2023!</b> Your team has been approved and you are ready to start the iGEM season!{% endblock %}
+{% block cover_img %}{% endblock %}
{% block page_content %}
diff --git a/wiki/pages/human-practices.html b/wiki/pages/human-practices.html
index 3eb92d80f7a63acd365fe4498eda3ef697098ee5..dc8456826599214e5816f024f03ac0f816820ecb 100644
--- a/wiki/pages/human-practices.html
+++ b/wiki/pages/human-practices.html
@@ -1,7 +1,7 @@
{% extends "layout.html" %}
{% block title %}Human Practices{% endblock %}
-{% block lead %}We ask every team to think deeply and creatively about whether their project is responsible and good for the world. Consider how the world affects your work and how your work affects the world.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/human-practices.jpg{% endblock %}
{% block page_content %}
diff --git a/wiki/pages/inclusivity.html b/wiki/pages/inclusivity.html
index e1b8add2da50839407ca27b430b42eb38bab9bff..1c8bbf1b340c6bfe47bff4e3ef08c7a5bd685c08 100644
--- a/wiki/pages/inclusivity.html
+++ b/wiki/pages/inclusivity.html
@@ -1,7 +1,7 @@
{% extends "layout.html" %}
{% block title %}Diversity and Inclusion{% endblock %}
-{% block lead %}Every individual, regardless of background or experience, should have an equal opportunity to engage with scientific knowledge and technological development.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/inclusivity.jpg{% endblock %}
{% block page_content %}
diff --git a/wiki/pages/measurement.html b/wiki/pages/measurement.html
index af066d8091d92cbb18956e57b357876001ed4604..3df626a45ccdb4728a183e775bcba25cc839fc18 100644
--- a/wiki/pages/measurement.html
+++ b/wiki/pages/measurement.html
@@ -1,7 +1,7 @@
{% extends "layout.html" %}
{% block title %}Measurement{% endblock %}
-{% block lead %}Synthetic Biology needs great measurement approaches for characterizing parts, and efficient new methods for characterizing many parts at once. Describe your measurement approaches on this page.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/measurement.jpg{% endblock %}
{% block page_content %}
diff --git a/wiki/pages/model.html b/wiki/pages/model.html
index cae119fdef6434118eb5657fcbc1e5c25aa11290..e4c417df78a9bf7ff799ccd3832fcb213e8910cc 100644
--- a/wiki/pages/model.html
+++ b/wiki/pages/model.html
@@ -1,7 +1,7 @@
{% extends "layout.html" %}
{% block title %}Model{% endblock %}
-{% block lead %}Explain your model's assumptions, data, parameters, and results in a way that anyone could understand.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/model.jpg{% endblock %}
{% block page_content %}
diff --git a/wiki/pages/notebook.html b/wiki/pages/notebook.html
index c955f7b10fc5e2133d130c370a1500c942063495..028c486c8f46d23d8fc7543b25ed357d548f50ba 100644
--- a/wiki/pages/notebook.html
+++ b/wiki/pages/notebook.html
@@ -1,34 +1,351 @@
{% extends "layout.html" %}
{% block title %}Notebook{% endblock %}
-{% block lead %}Document the dates you worked on your project. This should be a detailed account of the work done each day for your project.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/notebook.jpg{% endblock %}
+
+{% block detail_content1 %}{% endblock %}
+{% block detail_title_a1 %}Experimental preparation for week 1{% endblock %}
+{% block detail_content2 %}Experimental plan: Prepare the medium and experimental consumables and activate the plasmid-carrying strain
+Experimental procedure:
+1. Use of ultra-clean table: open the ultra-clean table, place the centrifuge tube plate, test tube rack, Marker pen, alcohol disinfection and sterilization, and conduct UV irradiation for 30 minutes.
+2. Sterilization: Prepare Eppendorf tubes, PCR tubes, and centrifuge tubes, and wrap them with gauze. Fill 2 boxes each of large, medium, small, and gun heads, using Velcro paper to wrap the shell, and using string/rubber band to bind the gun head box. Using pre-prepared LB medium, divide 20 tubes into 50 mL x 2 bottles of LB liquid medium. The above items were sterilized using 121 ° C for 15 min (this process was about 1.5 h). An experimental accident occurred during the filling of liquid medium: the medium was spilled on the ultra-clean table, and it was cleaned up properly afterwards.
+3. Activated strains: 1 tube of E. coli DH5α/ empty vector 1 and 1 tube of DH5α/ pheP gene. One tube of E. coli DH5α/ empty vector 2 and one tube of DH5α/ carrying PAL gene. Turn on the ultra-clean table, ventilated, put the sterilized LB test tube into the table, light the alcohol lamp, burn the test tube orifice and plug, put 100 μL of preserved bacteria solution into each test tube, add 2.5 μL Amp antibiotic mother solution, burn the test tube orifice and plug again. After sealing the tube, the tube was bundled and incubated on a shaker overnight (150 rpm, 37℃).
+
+{% endblock %}
+{% block detail_title_a2 %}2 weeks to extract the templates, began to construct the plasmid{% endblock %}
+{% block detail_content3 %}Experimental plan: Plasmid was extracted and the target fragment was amplified by PCR
+Experimental procedure:
+1. Plasmid extraction: Four kinds of plasmids DH5α/ empty vector 1 and one tube DH5α/ pheP gene were extracted according to the instruction of plasmid extraction kit. One tube of E. coli DH5α/ empty vector 2 and one tube of DH5α/ carrying PAL gene. The bacterial solution was centrifuged for 1min and then the precipitate was removed. p1 was added separately and resuspended, p2 was mixed upside down, p3 was mixed upside down, and centrifuged for 10min. 500μL BL reagent was added to the adsorption column, the waste solution was poured, the supernatant after centrifugation as described above was added, and the mixture was rinsed twice using pw rinse solution, and centrifuged for 1min each time. EB solution 50μL was added to the adsorption column membrane and centrifuged for 2min. The recovered DNA concentration was determined using Nanodrop and the DNA was stored at -20 ° C. In this step, most people successfully extracted the plasmid.
+{% endblock %}
+{% block detail_img1 %}https://static.igem.wiki/teams/5013/wiki/notebook/img1.png{% endblock %}
+{% block detail_content4 %}Strains and plasmid sample white floc after drip into the P3
+
+{% endblock %}
+{% block detail_img2 %}https://static.igem.wiki/teams/5013/wiki/notebook/img2.png{% endblock %}
+{% block detail_content5 %}
+The experimental work
+2. Dilute the primers
+The dry powder primer was centrifuged at 12,000 RPM for 1min, and the corresponding amount of distilled water was added to dilute the primer to the appropriate concentration
+{% endblock %}
+{% block detail_img3 %}https://static.igem.wiki/teams/5013/wiki/notebook/img3.jpg{% endblock %}
+{% block detail_content6 %}
+Centrifuge dry powder primer manipulation
+
+3.PCR reaction:
+(1) Target genes pheP, PAL and two kinds of vector DNA fragments were amplified by high fidelity enzyme Pfu. The reaction system is as follows:
+Pfu 0.5 μL
+10xPfu buffer 5 μL
+Primer 1 1 μL
+Primer 2 1 μL
+Template (plasmid extracted above) : 1μL
+25μL dNTP Mixture
+20 μL of sterilized water
+(2) Perform the PCR amplification procedure
+At 98 ° C for 5 min
+The following 3-step reaction was performed for 30 cycles
+98℃ 10s
+55℃ 5s
+72℃ 1min/kb
+72℃ 8 min
+The plasmids, primers and products required for PCR are shown in the table below:
+
+{% endblock %}
+{% block detail_title_a3 %}3 weeks agarose gel electrophoresis, plastic recycling, double enzyme, recycling{% endblock %}
+{% block detail_content7 %}Experimental plan: Agarose gel electrophoresis, gel recovery, double enzyme digestion, recovery
+Experimental procedure:
+1 Agarose gel electrophoresis: The above PCR products were added to a previously prepared agarose gel with 5μL DL 5000 DNA marker and subjected to electrophoresis (120V, 20 min). At the end of electrophoresis, the gel imaging system was used for observation. The target DNA gel was cut, placed in a 2 mL centrifuge tube, and labeled.
+{% endblock %}
+{% block detail_img4 %}https://static.igem.wiki/teams/5013/wiki/notebook/img4.jpg{% endblock %}
+{% block detail_content8 %}
+Plot of gel electrophoresis results
+2. Gel recovery: The PCR product DNA gel was recovered according to the gel recovery kit instructions
+Add 500 microliters of equilibrium solution BL to the adsorption column CA2 and centrifuge for 1 min, pour out the waste liquid in the collection tube, and put the adsorption column back into the collection tube. A single band of target DNA was cut from the agarose gel into a clean centrifuge tube. Add an equal volume of solution PN to the gel block and place in a water bath at 50 ° C until the gel block is completely dissolved. The solution obtained in the previous step was added to another adsorption column CA2, placed at room temperature for 2 min, and centrifuged for 1min. Add 600 microliters of bleach solution PW to the adsorption column CA2 (please check whether absolute ethanol has been added before use), centrifuge for 1min, repeat rinsing twice, and let dry. Add an appropriate amount of elution buffer EB to the middle position of the adsorption membrane and place it at room temperature for 2 min. The DNA solution was collected by centrifugation for 2min. The recovered DNA concentration was determined using Nanodrop.
+{% endblock %}
+{% block detail_img5 %}https://static.igem.wiki/teams/5013/wiki/notebook/img5.png{% endblock %}
+{% block detail_content9 %}
+Completed rubber recycling product concentration determination of DNA
+3. Double digestion: EcoRI and XhoI were used to double digest the extracted plasmid pET23b and the amplified target DNA fragment. 5x20 μL reaction. The 20 μL digestion system is as follows:
+1 μg of DNA
+1 μL of EcoRI
+1 μL XhoI
+4 μL 10xY buffer
+H2O up to 20μL
+The enzyme was digested at 37 ° C for 4h
+4 Agarose gel electrophoresis: The digested product was added to the agarose gel, 5μL of DL 5000 DNA marker was added, and electrophoresis was performed (120V, 20 min). At the end of electrophoresis, the gel imaging system was used for observation. The target DNA gel was cut, placed in a 2 mL centrifuge tube, and labeled. The product was extracted successfully, but the amount was small. After that, the second gum recovery was carried out.
+{% endblock %}
+{% block detail_img6 %}https://static.igem.wiki/teams/5013/wiki/notebook/img6.png{% endblock %}
+{% block detail_content10 %}
+The researchers cut gel
+{% endblock %}
+{% block detail_img7 %}https://static.igem.wiki/teams/5013/wiki/notebook/img7.png{% endblock %}
+{% block detail_content11 %}
+{% endblock %}
+{% block detail_img8 %}https://static.igem.wiki/teams/5013/wiki/notebook/img8.png{% endblock %}
+{% block detail_content12 %}
+Photo of the experimenter at work
+
+{% endblock %}
+{% block detail_title_a4 %}Competent cells were prepared at week 4{% endblock %}
+{% block detail_content13 %}Experimental plan: E. coli DH5α and Rosetta were activated to prepare competent cells
+Experimental procedure:
+1. Activated strains: including Escherichia coli DH5α and Rosetta. Turn on the ultra-clean table, ventilate, put the sterilized LB test tube into the table, light the alcohol lamp, burn the test tube mouth and test tube stopper, connect 100 μL of preserved bacteria solution into each test tube, and burn the test tube mouth and test tube stopper again. The test tube was closed, bundled, and incubated on a shaker overnight (220 rpm, 37℃).
+Experimental procedure:
+2. Preparation of competent cells:
+(1) Open the ultra-clean stage, put in 1 mL tip, 200 μL tip, 5 mL tip, 2mL and 30 mL centrifuge tubes, and 2 x 50 mL LB medium for UV irradiation. The 0.1 M cacl 2 and 15% 0.1 M cacl 2 solution were pre-cooled in a refrigerator at 4 ° C.
+(2) DH5α and Rosetta competent cells were prepared as follows: pre-activated and packed DH5α and Rosetta bacteria were precooled in a centrifuge at 4 ° C, centrifuged at 5000 g for 10 min, the supernatant was discard, and 800 microliters of 0.1 M CaCl2 solution was added to an ice bath for 40 min. After centrifugation again (4000 g, 10 min), the supernatant was discarded, and 100 microliters of 15% glycerol 0.1 M CaCl2 were added to each tube to resuspend the bacteria. Aliquots of competent cells into 2 mL centrifuge tubes of 100μL per tube were dispensed on ice. After labeling, the competent cells were stored in the refrigerator at -80℃.
+{% endblock %}
+{% block detail_img9 %}https://static.igem.wiki/teams/5013/wiki/notebook/img9.png{% endblock %}
+{% block detail_content14 %}
+{% endblock %}
+{% block detail_img10 %}https://static.igem.wiki/teams/5013/wiki/notebook/img10.png{% endblock %}
+{% block detail_content15 %}
+1. Pre-activated aliquots of DH5α and Rosetta bacterial samples
+2. Bacterial samples in an ice bath
+3 Add the CaCl2 solution
+4. After secondary centrifugation of the bacterial sample, white precipitate was produced
+5 Add glycerol CaCl2 mixing buffer
+
+{% endblock %}
+{% block detail_title_a5 %}Week 5 Connect and transform{% endblock %}
+{% block detail_content16 %}Experimental plan: Prepare solid AGAR plates containing antibiotics, perform ligation reactions, and transform ligation products
+Experimental procedure:
+1. Preparation of antibiotic plates: LB solid medium (4.5 g AGAR added to 300 mL LB liquid medium) was prepared and sterilized at 121 ° C for 15 min by autoclave. After cooling to about 60℃, add 150 μL of 100mg/mL Amp antibiotic, shake evenly, and pour into the disposable Petri dish (about 10-15 plates).
+{% endblock %}
+{% block detail_img11 %}https://static.igem.wiki/teams/5013/wiki/notebook/img11.jpg{% endblock %}
+{% block detail_content17 %}
+Pour the liquid LB medium into the disposable Petri dish
+2 Ligation reaction: The vector was ligated by double enzyme digestion of the target fragment. The ligation system is as follows (Note: This system is an empirical system, and the optimal linking vector/target fragment volume ratio can also be calculated according to the instructions) :
+T4 DNA ligase 1 μL
+2 μL of 10 x ligation buffer
+ 1 μL of Linear plasmid
+ Insert 6 μL of DNA
+ Connections were made using a PCR apparatus at 16 ° C for 1h. pET23b-insert was obtained, and the ligation product was stored at -20 ° C.
+3. Transformation of ligation products: 10 μL of the above ligation products were added to 100 μL competent cells, followed by an ice bath for 30 min, a water bath at 42 ° C for 60 s, and an ice bath for 5 min. Then, 1 mL LB liquid medium was added and incubated for 1 h for recovery (37 ° C, 150 rpm). In an ultra-clean stage, 100 μL of recovery solution was taken and the Amp antibiotic plate was coated with a sterile coating rod.
+
+{% endblock %}
+{% block detail_title_a6 %}At the 6th week, the two successfully constructed plasmids were extracted and expanded{% endblock %}
+{% block detail_content18 %}Experimental plan: colony PCR verification, expansion of positive clones, extraction and verification of the correct plasmid and transformation
+Experimental procedure:
+1.PCR verification: Six colonies were labeled on each plate. Half of the colonies were picked and used as templates for colony PCR using Taq enzyme. The primers are TF, TR. The reaction system and procedures were carried out according to the instructions. Configure a 50 mL agarose gel, using DL 2000 DNA marker, and perform electrophoresis. The correct size of the band indicates that the plasmid was successfully constructed
+2. Expanded culture: The correct colonies were selected and added to 5 mL LB liquid medium, and 2.5 μL Amp antibiotic was added to expand the culture.
+3. Plasmid extraction: Two kinds of plasmids were extracted according to the instruction of plasmid extraction kit. The bacterial solution was centrifuged for 1min and then the precipitate was taken. p1 was added separately and resuspended, p2 was mixed upside down, p3 was mixed upside down, and centrifuged for 10min. 500μL BL reagent was added to the adsorption column, the waste solution was poured, the supernatant after centrifugation as described above was added, and the mixture was rinsed twice using pw rinse solution, and centrifuged for 1min each time. EB solution 50μL was added to the adsorption column membrane and centrifuged for 2min. The recovered DNA concentration was determined using Nanodrop and the DNA was stored at -20 ° C. In this step, most people successfully extracted the plasmid.
+4. Transformation: Verify the correct plasmid 1 transformation into expression Escherichia coli.
+
+{% endblock %}
+{% block detail_title_a7 %}The ability of cells to metabolize phe was measured at the 7th week{% endblock %}
+{% block detail_content19 %}Experimental plan:
+Experimental procedure:
+1 Incubation, ELISA: To measure the capacity of whole cells to metabolization of phe, the PAL engineered strain was resuspended to an OD600 of 0.1 in 1 mL of Phe assay buffer (M9 0.5% glucose containing 1 mM millimoles per liter of Phe) and incubated in microtubes. The amount of Phe was then determined according to the phenylalanine ELISA kit. , the results are shown in FIG. A below.
+2. The strains were resuspended in 1 mL Phe experiment buffer (M9 0.5% glucose containing 1 mM Phe) to an OD600 of 0.1, placed in microtubes, and incubated at 37°C for 1h. TCA concentration was determined with OD290 as shown in FIG. B.
+3 Simultaneously determine the effect of pH on PAL enzyme activity, as shown in Figure C.
+
+{% endblock %}
+{% block detail_img12 %}https://static.igem.wiki/teams/5013/wiki/notebook/img12.png{% endblock %}
+{% block detail_content20 %}
+{% endblock %}
+{% block detail_title_a8 %}At week 8, the effect of the hypoxic promoter was tested to verify whether the hypoxic promoter could effectively activate PAL expression{% endblock %}
+{% block detail_content21 %}Experimental plan: To test the effect of the hypoxic promoter and PAL expression
+Experimental procedure:
+1. The tested engineering bacteria were statically cultured in a CO2 incubator (adjust the O2 concentration to 0%, 10% and 20%). After 16h of growth, the absorbance (OD600) and mRFP fluorescence intensity data were measured using a microplate reader.
+2. The tested engineered bacteria were statically cultured in a CO2 incubator (O2 concentration was adjusted to 0%, 10% and 20%). The strains were resuspended in 1 mL Phe experiment buffer (M9 0.5% glucose containing 1 mM Phe) to an OD600 of 0.1, placed in microtubes, and incubated at 37°C for 1h. TCA concentration was determined using OD290. The Phe content was measured using an Elisa kit.
+{% endblock %}
+{% block detail_img13 %}https://static.igem.wiki/teams/5013/wiki/notebook/img13.png{% endblock %}
+{% block detail_content22 %}
+
+{% endblock %}
+{% block detail_title_a9 %}Trans-cinnamic acid (TCA) promoter test and determination of PAL activity at week 9{% endblock %}
+{% block detail_content23 %}Experimental plan: Promoter testing and determination of PAL activity
+Experimental procedure:
+1 The strain was resuspended in 1 mM Phe experiment buffer to an OD600 of 0.1 and placed in a microtube, and an additional 1 mM TCA was added to the test group. The cells were incubated at 37°C for 1h. Phe content was determined according to the phenylalanine ELISA kit, and TCA content was tested using OD290.
+
+{% endblock %}
+{% block detail_title_a10 %}At the 10th week, the signal amplification switch of TP901 was coupled to test the fluorescent protein expression under different TCA content{% endblock %}
+{% block detail_content24 %}Experimental plan: The signal amplification switch of TP901 was coupled to test the fluorescent protein expression
+Experimental procedure:
+1. The same procedure as above was used to construct the plasmid, mRFP gene was coupled to the downstream of TCA promoter pSenCA, and the fluorescent protein expression was measured under different TCA content.
+2. Fluorescence microscope verification experiment:
+(1) Sample preparation: appropriate amount (about 30 μl) of wild type Rosetta and Rosetta-engineered bacteria droplets were dropped onto the slide and covered with a cover slip.
+(2) Turn on the switch of the power control box of the high-pressure mercury lamp.
+(3) Insert the light stopper to interrupt the light path.
+(4) Preheat for 5-10 min.
+(5) Place the slide with the sample on the stage.
+(6) Select the objective lens (in order of first low power, then high power).
+(7) Rotate the turntable of the spectroscope assembly to select the spectroscope assembly for observing mRFP (red fluorescence).
+(8) Adjust the focal length through thick and thin screws.
+(9) Open the computer connected to the microscope and click on the digital imaging system software to collect digital images.
+3. Microplate reader was used to test fluorescent protein expression: 100 μL of wild-type Rosetta, recombinant strain solution was taken in an ultra-clean stage. The supernatant was added to a 96-well plate for 4-6 replicates. The detection conditions of the microplate reader were set (mRFP excitation λ: 584 nm/10 nm, emission λ: 607/10 nm), and the readings were taken. The data were recorded using Excel.
+
+
+
+
+{% endblock %}
{% block page_content %}
-<div class="row mt-4">
- <div class="col-lg-8">
- <h2>What should this page contain?</h2>
- <hr>
- <ul>
- <li>Chronological notes of what your team is doing.</li>
- <li>Brief descriptions of daily important events.</li>
- <li>Pictures of your progress.</li>
- <li>Mention who participated in what task.</li>
- </ul>
+<div class="page-content flex-row">
+ <img class="bgImage" src="https://static.igem.wiki/teams/5013/wiki/desc/desc-bg.jpg">
+
+ <!-- 侧边æ -->
+ <div class="sidebar flex-column">
+
+ <!-- todo -->
+ <a class="sidebar-a" href="#a1">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a1()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a2">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a2()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a3">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a3()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a4">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a4()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a5">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a5()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a6">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a6()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a7">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a7()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a8">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a8()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a9">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a9()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a10">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a10()}}</div>
+ </div>
+ </a>
+
</div>
- <div class="col-lg-4">
- <h2>Inspirations</h2>
- <hr>
- <ul>
- <li><a href="http://2018.igem.org/Team:Munich/Notebook">2018 Munich</a></li>
- <li><a href="https://2019.igem.org/Team:Georgia_State/Notebook">2019 Georgia State</a></li>
- <li><a href="https://2019.igem.org/Team:Newcastle/Notebook">2019 Newcastle</a></li>
- <li><a href="https://2020.igem.org/Team:IISER-Pune-India/Notebook">2020 IISER Pune India</a></li>
- <li><a href="https://2020.igem.org/Team:Lund/Notebook">2020 Lund</a></li>
- <li><a href="https://2020.igem.org/Team:NOVA_LxPortugal/Notebook">2020 NOVA LxPortugal</a></li>
- <li><a href="https://2020.igem.org/Team:RDFZ-China/NoteBook">2020 RDFZ China</a></li>
- </ul>
+
+ <!-- 文案æ£æ–‡ -->
+ <div class="desc-content flex-column">
+ <!-- todo -->
+ <pre class="desc-content-common-text">{{self.detail_content1()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a1">{{self.detail_title_a1()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content2()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a2">{{self.detail_title_a2()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content3()}}</pre>
+ <img loading="lazy" src="{{self.detail_img1()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content4()}}</pre>
+ <img loading="lazy" src="{{self.detail_img2()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content5()}}</pre>
+ <img loading="lazy" src="{{self.detail_img3()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content6()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a3">{{self.detail_title_a3()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content7()}}</pre>
+ <img loading="lazy" src="{{self.detail_img4()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content8()}}</pre>
+ <img loading="lazy" src="{{self.detail_img5()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content9()}}</pre>
+ <img loading="lazy" src="{{self.detail_img6()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content10()}}</pre>
+ <img loading="lazy" src="{{self.detail_img7()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content11()}}</pre>
+ <img loading="lazy" src="{{self.detail_img8()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content12()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a4">{{self.detail_title_a4()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content13()}}</pre>
+ <img loading="lazy" src="{{self.detail_img9()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content14()}}</pre>
+ <img loading="lazy" src="{{self.detail_img10()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content15()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a5">{{self.detail_title_a5()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content16()}}</pre>
+ <img loading="lazy" src="{{self.detail_img11()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content17()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a6">{{self.detail_title_a6()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content18()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a7">{{self.detail_title_a7()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content19()}}</pre>
+ <img loading="lazy" src="{{self.detail_img12()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content20()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a8">{{self.detail_title_a8()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content21()}}</pre>
+ <img loading="lazy" src="{{self.detail_img13()}}" />
+ <pre class="desc-content-common-text">{{self.detail_content22()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a9">{{self.detail_title_a9()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content23()}}</pre>
+ <div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a10">{{self.detail_title_a10()}}</div>
+ </div>
+ <pre class="desc-content-common-text">{{self.detail_content24()}}</pre>
+
</div>
+
</div>
+
{% endblock %}
diff --git a/wiki/pages/plant.html b/wiki/pages/plant.html
index 7a77bef5ca6907e6e3ef7e57b1da6097017b04db..1900281667d0ca3618dea6e09422db2d4207f35a 100644
--- a/wiki/pages/plant.html
+++ b/wiki/pages/plant.html
@@ -1,7 +1,7 @@
{% extends "layout.html" %}
{% block title %}Plant{% endblock %}
-{% block lead %}This award is designed to celebrate exemplary work done in plant synthetic biology.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/plant.jpg{% endblock %}
{% block page_content %}
diff --git a/wiki/pages/proof-of-concept.html b/wiki/pages/proof-of-concept.html
new file mode 100644
index 0000000000000000000000000000000000000000..1124a25f052c90783877835c5c7001751752428c
--- /dev/null
+++ b/wiki/pages/proof-of-concept.html
@@ -0,0 +1,103 @@
+{% extends "layout.html" %}
+
+{% block title %}Proof of Concept{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/proof-of-concept.jpg{% endblock %}
+
+{% block detail_content1 %}{% endblock %}
+{% block detail_img1 %}https://static.igem.wiki/teams/5013/wiki/proof/img1.png{% endblock %}
+{% block detail_content2 %}<div class="desc-content-img-desc">Figure 1 Schematic representation of the overall principle of PAL gene expression.</div>{% endblock %}
+{% block detail_title_a1 %}Construct an engineered strain with anaerobic promoter pPep to drive the expression of Pal.{% endblock %}
+{% block detail_content3 %}Phenylalanine ammonia-lyase (PAL) is an important enzyme that catalyzes the deamination of phenylalanine (Phe), removing the amino group that cannot be processed by PKU patients.The principle is illustrated in Figure 2G.Initially, we used a constitutive promoter to express the pal gene and measured Phe levels using a phenylalanine ELISA kit, while TCA concentration was determined by measuring OD290 with an enzyme immunoassay analyzer. The results, as shown in Figure 2A and 2B, demonstrate that after PAL expression, Phe levels decreased significantly, while TCA levels increased, indicating that PAL can effectively degrade Phe into TCA. Considering the role of PAL in anaerobic environments, we decided to test its expression using an anaerobic promoter, and the results, as shown in Figure 2F, indicated normal expression under anaerobic conditions. Similarly, Phe and TCA levels were measured, as shown in Figure 2C and 2D. When oxygen concentration was lower, TCA production was higher, indicating that PAL can play a significant role under anaerobic conditions. Furthermore, we validated the effect of environmental pH on PAL metabolic capacity, as shown in Figure 2E. Within the range of pH 5 to 8, PAL activity increased with increasing pH, with pH 8 being the optimal pH.
+
+{% endblock %}
+{% block detail_img2 %}https://static.igem.wiki/teams/5013/wiki/proof/img2.png{% endblock %}
+{% block detail_content4 %}<div class="desc-content-img-desc">Figure 2 Hypoxia-induced promoter pPep drives PAL expression.</div>
+
+{% endblock %}
+{% block detail_title_a2 %}Utilizing the trans-cinnamic acid biosensor pSenCA and signal amplification switch for efficient gene expression.{% endblock %}
+{% block detail_content5 %}We designed a trans-cinnamic acid (TCA) promoter and TP901 amplification signal switch system to efficiently express PAL in an in vitro transformation system. The schematic diagram of the trans-cinnamic acid (TCA) promoter is shown in Figure 3A, and the schematic diagram of the trans-cinnamic acid (TCA) promoter and TP901 amplification signal switch system is shown in Figure 3B. Firstly, we constructed mRFP downstream of the pTCA promoter and detected its expression, as shown in Figure 3C. After adding 1mM TCA, there was a significant increase in mRFP fluorescence intensity. Secondly, we measured the content of Phe and TCA in the engineered strain using the same method, as shown in Figure 3D and E. After adding 1mM TCA, the Phe content decreased from around 1mM to around 0mM, while the TCA content increased from 0mM to around 2mM. Therefore, pTCA has a good activation ability for PAL expression.
+ To verify if the TP901 amplification signal switch system can promote gene expression downstream of pSenCA TCA promoter, we coupled mRFP downstream of pSenCA and added the TP901 amplification signal switch system. We tested the expression of fluorescent protein at different TCA concentrations. From Figure 3F, it can be observed that in an environment with 1mM TCA, the strain with the TP901 signal amplification switch system added downstream of pTCA had a fluorescence intensity of around 40A.U, which was significantly higher compared to the control group. This indicates that TP901 has a significant effect in promoting downstream gene expression.
+
+
+{% endblock %}
+{% block detail_img3 %}https://static.igem.wiki/teams/5013/wiki/proof/img3.png{% endblock %}
+{% block detail_content6 %}<div class="desc-content-img-desc">Figure3 Utilizing the trans-cinnamic acid biosensor pSenCA and signal amplification switch for efficient gene expression.</div>
+{% endblock %}
+{% block detail_title_a3 %}References{% endblock %}
+{% block detail_content7 %}1. Chien, Tiffany, et al. "Enhancing the tropism of bacteria via genetically programmed biosensors." Nature biomedical engineering 6.1 (2022): 94-104.
+2. Courbet, Alexis, et al. "Detection of pathological biomarkers in human clinical samples via amplifying genetic switches and logic gates." Science translational medicine 7.289 (2015): 289ra83-289ra83.
+3. Flachbart, Lion Konstantin, Sascha Sokolowsky, and Jan Marienhagen. "Displaced by deceivers: prevention of biosensor cross-talk is pivotal for successful biosensor-based high-throughput screening campaigns." ACS synthetic biology 8.8 (2019): 1847-1857.
+4. Binder, Stephan, et al. "A high-throughput approach to identify genomic variants of bacterial metabolite producers at the single-cell level." Genome biology 13.5 (2012): 1-12.
+5. Pi, Jing, PETER J. Wookey, and A. J. Pittard. "Cloning and sequencing of the pheP gene, which encodes the phenylalanine-specific transport system of Escherichia coli." Journal of bacteriology 173.12 (1991): 3622-3629.
+6. Isabella, Vincent M., et al. "Development of a synthetic live bacterial therapeutic for the human metabolic disease phenylketonuria." Nature biotechnology 36.9 (2018): 857-864.
+
+
+
+
+{% endblock %}
+
+{% block page_content %}
+
+<div class="page-content flex-row">
+ <img class="bgImage" src="https://static.igem.wiki/teams/5013/wiki/desc/desc-bg.jpg">
+
+ <!-- 侧边æ -->
+ <div class="sidebar flex-column">
+
+ <!-- todo -->
+ <a class="sidebar-a" href="#a1">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a1()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a2">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a2()}}</div>
+ </div>
+ </a>
+<a class="sidebar-a" href="#a3">
+ <div class="sidebar-item flex-row align-center">
+ <div class="sidebar-item-circle">
+ </div>
+ <div class="sidebar-item-text">{{self.detail_title_a3()}}</div>
+ </div>
+ </a>
+
+ </div>
+
+ <!-- 文案æ£æ–‡ -->
+ <div class="desc-content flex-column">
+ <!-- todo -->
+ <pre class="desc-content-common-text">{{self.detail_content1()}}</pre>
+<img loading="lazy" src="{{self.detail_img1()}}" />
+<pre class="desc-content-common-text">{{self.detail_content2()}}</pre>
+<div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a1">{{self.detail_title_a1()}}</div>
+ </div>
+<pre class="desc-content-common-text">{{self.detail_content3()}}</pre>
+<img loading="lazy" src="{{self.detail_img2()}}" />
+<pre class="desc-content-common-text">{{self.detail_content4()}}</pre>
+<div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a2">{{self.detail_title_a2()}}</div>
+ </div>
+<pre class="desc-content-common-text">{{self.detail_content5()}}</pre>
+<img loading="lazy" src="{{self.detail_img3()}}" />
+<pre class="desc-content-common-text">{{self.detail_content6()}}</pre>
+<div class="desc-content-title flex-row align-center">
+ <div class="desc-content-title-circle"></div>
+ <div class="desc-content-title-text" id="a3">{{self.detail_title_a3()}}</div>
+ </div>
+<pre class="desc-content-common-text">{{self.detail_content7()}}</pre>
+
+ </div>
+
+ </div>
+
+
+{% endblock %}
\ No newline at end of file
diff --git a/wiki/pages/results.html b/wiki/pages/results.html
index fae8f05c86f098fff4718f9bf385da5b00ad2189..522558149f981610f73a371dc26a1f9f51edf794 100644
--- a/wiki/pages/results.html
+++ b/wiki/pages/results.html
@@ -1,7 +1,7 @@
{% extends "layout.html" %}
{% block title %}Results{% endblock %}
-{% block lead %}You can describe the results of your project and your future plans here.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/results.jpg{% endblock %}
{% block page_content %}
diff --git a/wiki/pages/safety.html b/wiki/pages/safety.html
index 3e7d1ed3bd3ae09c809b027dc4dfe75c17a866e4..4a1a95db29b8214713fc16107bd60d7500089944 100644
--- a/wiki/pages/safety.html
+++ b/wiki/pages/safety.html
@@ -1,7 +1,7 @@
{% extends "layout.html" %}
{% block title %}Safety{% endblock %}
-{% block lead %}Describe all the safety issues of your project.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/safety.jpg{% endblock %}
{% block page_content %}
diff --git a/wiki/pages/software.html b/wiki/pages/software.html
index a4e352b4e1eb89e04acb4c340cad169ae0961ae4..d03d022817a5ec6d09da5a2596098d0442dda204 100644
--- a/wiki/pages/software.html
+++ b/wiki/pages/software.html
@@ -1,7 +1,7 @@
{% extends "layout.html" %}
{% block title %}Software{% endblock %}
-{% block lead %}Software in iGEM should make synthetic biology based on standard parts easier, faster, better or more accessible to our community.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/software.jpg{% endblock %}
{% block page_content %}
diff --git a/wiki/pages/sustainable.html b/wiki/pages/sustainable.html
index e209db3663c566d44ae24d76f529c8af364ebdae..bce1e124d527085af0adfce0ce7d6af0c9117968 100644
--- a/wiki/pages/sustainable.html
+++ b/wiki/pages/sustainable.html
@@ -1,7 +1,7 @@
{% extends "layout.html" %}
{% block title %}Sustainable Development Goals{% endblock %}
-{% block lead %}Describe how you have evaluated your project ideas against one or more of the SDGs.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/sustainable.jpg{% endblock %}
{% block page_content %}
diff --git a/wiki/pages/team.html b/wiki/pages/team.html
index 1045da132b35c96d85b02701e2f71b74dd6398c1..5c3d9c5315432e924bc6bd7a6f7bca5c0c5b190b 100644
--- a/wiki/pages/team.html
+++ b/wiki/pages/team.html
@@ -1,7 +1,9 @@
{% extends "layout.html" %}
{% block title %}Team{% endblock %}
-{% block lead %}On this page you can introduce your team members, instructors, and advisors.{% endblock %}
+{% block cover_img %}https://static.igem.wiki/teams/5013/wiki/cover/team.jpg{% endblock %}
+
+
{% block page_content %}