From a7bbce07e59afe235606ce16a827c720ad8f9a7d Mon Sep 17 00:00:00 2001
From: HouTeng Chan <ht-chen21@mails.tsinghua.edu.cn>
Date: Tue, 1 Oct 2024 12:02:31 +0000
Subject: [PATCH] Update file colonization.html
---
wiki/pages/colonization.html | 156 +++++++++++++++++++----------------
1 file changed, 83 insertions(+), 73 deletions(-)
diff --git a/wiki/pages/colonization.html b/wiki/pages/colonization.html
index a3cd4e7f..960d4382 100644
--- a/wiki/pages/colonization.html
+++ b/wiki/pages/colonization.html
@@ -37,7 +37,6 @@
<div class="sidebar">
<ul>
<li><a href="#Design">Design</a></li>
- <li><a href="#Experiment">Experiment</a></li>
<li><a href="#Result">Result</a></li>
<li><a href="#Protocol">Protocol</a></li>
<li><a href="#Notebook">Notebook</a></li>
@@ -57,59 +56,78 @@
<h2 id="Design">
<h2>Design</h2>
<hr>
- <p>In order to enable our engineered bacteria to specifically function at the intestinal lesions of IBD patients, thereby achieving better and more precise therapeutic effects, we designed a colonization system in Saccharomyces cerevisiae. The system consists of two components: the tetrathionate sensor TtrSR and the adhesive protein Als3.</p>
+ <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
+ <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/pic1.jpg" alt="ibd_figure" class="shadowed-image" style="width: 60%; max-width: 600px;">
+ <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 1 Schematic Representation of colonization Experimental Design</p>
+ </div>
+ <p>To enable our engineered bacteria to specifically function at the small intestinal lesions in IBD patients, we designed the colonization system. This system consists of two main components: one is the tetrathionate sensor TtrSR, and the other is the adhesion protein Als3. TtrSR is a two-component system from the marine bacterium Shewanella halifaxensis HAW-EB4, which can detect extracellular tetrathionate chemical signals and promote the expression of downstream genes in the signaling pathway. Als3 is a cell surface protein from Candida albicans, which acts as an adhesin, mediating adhesion to epithelial cells, endothelial cells, and extracellular matrix proteins. We chose Saccharomyces cerevisiae as the chassis organism for engineering the bacteria. By expressing the TtrSR system and Als3 protein in Saccharomyces cerevisiae, we will achieve specific colonization at the small intestinal lesions in IBD patients.</p>
+ <p>Our experimental design is divided into two parts: verify the functionality of the TtrSR system in Saccharomyces cerevisiae and test the adhesion ability of Saccharomyces cerevisiae after expressing Als3. We independently validated each component of the colonization system in the engineered Saccharomyces cerevisiae and ultimately plan to integrate them together.</p>
</div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
- <h3>Tetrathionate sensor TtrSR</h3>
- <p>To locate the site of the lesion, we try to find a molecule which can specifically characterize IBD. Current research suggests that thiosulfate and tetrathionate can serve as indicators of intestinal inflammation (Levitt et al., 1999), and the levels of thiosulfate or tetrathionate are directly proportional to the degree of intestinal inflammation. A recent study constructed a tetrathionate sensor in E. coli (Kristina N-M Daeffler, 2017)</p>
- </div>
- <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
- <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/ttrs.png" alt="ibd_figure" class="shadowed-image" style="width: 50%; max-width: 450px;">
- <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 1 Schematic of the tetrathionate sensor TtrSR</p>
+ <h3>Colonization system</h3>
+ <p>Our colonization system is primarily composed of two parts:</p>
+ <p>Input signal - Tetrathionate sensor TtrR</p>
+ <p>Signal pathway - TtrRS two-component system</p>
+ <p>Output signal - Als3 adhesion protein</p>
+ <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
+ <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/pic2.png" alt="ibd_figure" class="shadowed-image" style="width: 60%; max-width: 600px;">
+ <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 2 Schematic Diagram of colonization System</p>
+ </div>
</div>
- <p>This system is the TtrSR two-component system (TCS) from the marine bacterium Shewanella halifaxensis HAW-EB4. The TCS consists of TtrS and TtrR. TtrS is a membrane-bound sensor histidine kinase (SK), which can phosphorylate the cytoplasmic response regulator (RR) TtrR in the presence of tetrathionate. Phosphorylated TtrR activates the expression of downstream genes through the ttrB promoter (PttrB).</p>
</div>
<div class="row mt-4">
<div class="col-lg-12">
- <h3>Adhesive protein Als3</h3>
- <p>Agglutinin-like sequence protein 3 (Als3) is a cell surface glycoprotein of Candida albicans that plays a significant role in vitro adhesion and biofilm formation. Als3 is essential for the binding of Candida albicans hyphae to various host cell surface receptor proteins, and it induces endocytosis by binding to E-cadherin on epithelial cells (Wang Tianming et al., Candida-epithelial interactions, 2023). Since our chassis organism, Saccharomyces cerevisiae, is also a fungus, similar to Candida albicans, expressing Als3 on the surface of S. cerevisiae cells could potentially enable binding to E-cadherin on intestinal epithelial cells, thereby allowing the engineered bacteria to colonize the small intestine.</p>
- <p>In the colonization system, we designed to express TtrSR along with Als3 in Saccharomyces cerevisiae. Als3 is located downstream of the ttrB promoter. After expressing this system in S. cerevisiae, when tetrathionate is present in the intestine (at the site of IBD lesions), the tetrathionate sensor is activated, leading to the expression of the Als3 protein. This ultimately enables the engineered bacteria to adhere to intestinal epithelial cells, thereby achieving colonization.</p>
- </div>
- <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
- <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/" alt="ibd_figure" class="shadowed-image" style="width: 90%; max-width: 800px;">
- <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 2 Schematic of the colonization system</p>
+ <h3>Tetrathionate Sensor TtrSR</h3>
+ <p>To identify the lesion site, we need a molecule that specifically characterizes IBD. Current research suggests that thiosulfate and tetrathionate can serve as indicators of intestinal inflammation (Levitt et al., 1999), and the levels of thiosulfate or tetrathionate are directly proportional to the severity of intestinal inflammation. A previous study constructed a tetrathionate sensor in E. coli (Kristina N-M Daeffler, 2017). This system is the TtrSR two-component system (TCS) from the marine bacterium Shewanella halifaxensis HAW-EB4. The TCS includes TtrS, a membrane-bound sensor histidine kinase (SK), which can phosphorylate the cytoplasmic response regulator (RR) TtrR in the presence of tetrathionate. Phosphorylated TtrR activates the expression of downstream genes through the ttrB promoter (PttrB).</p>
+ <p>To verify the effectiveness of the tetrathionate sensor TtrSR, we designed the corresponding plasmids. We planed to express EGFP downstream of ttrB and validate the effect of TtrSR by testing the fluorescence intensity of the cells. We used Ura-HIS nutrient-deficient medium to select yeast that had been successfully transformed. Then, we tested the effectiveness of the TtrSR system using confocal microscopy. For more details, please refer to protocol.</p>
+ <p>Aim:</p>
+ <p>To confirm the effectiveness of TtrRS in Saccharomyces cerevisiae</p>
+ <p>Constructs:</p>
+ <p>pYES2-SV40-ttrS</p>
+ <p>pESC-SV40-ttrR-PttrB-EGFP</p>
+ <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
+ <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/pic3.png" alt="ibd_figure" class="shadowed-image" style="width: 40%; max-width: 400px;">
+ <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 3 pYES2-SV40-ttrS plasmid</p>
+ </div>
+ <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
+ <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/pic4.png" alt="ibd_figure" class="shadowed-image" style="width: 40%; max-width: 400px;">
+ <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 4 pESC-SV40-ttrR-PttrB-EGFP plasmid</p>
+ </div>
+ <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
+ <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/pic5.jpg" alt="ibd_figure" class="shadowed-image" style="width: 40%; max-width: 400px;">
+ <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 5 result of transformation</p>
+ </div>
</div>
- <p>This system is the TtrSR two-component system (TCS) from the marine bacterium Shewanella halifaxensis HAW-EB4. The TCS consists of TtrS and TtrR. TtrS is a membrane-bound sensor histidine kinase (SK), which can phosphorylate the cytoplasmic response regulator (RR) TtrR in the presence of tetrathionate. Phosphorylated TtrR activates the expression of downstream genes through the ttrB promoter (PttrB).</p>
</div>
<div class="row mt-4">
<div class="col-lg-12">
- <h2 id="Experiment">
- <h2>Experiment</h2>
- <hr>
- <p>To test the effectiveness of the tetrathionate sensor TtrSR, we designed the corresponding plasmids (Fig.3 ttrR, Fig.4 ttrS). We arranged for EGFP to be expressed downstream of ttrB and validated the effectiveness of TtrSR by measuring the fluorescence intensity of the cells. After successfully transforming the plasmids into Saccharomyces cerevisiae, we selected three well-growing colonies for amplification. （Fig. after transforming）Then, we divided the cells from the same clone into two groups, one group induced with 1mM tetrathionate and the other group without inducer as control group. After 12 hours of induction, an appropriate amount of bacterial solution was prepared into temporary slides and the fluorescence intensity was tested using a fluorescence confocal microscope. For more details, please refer to the protocol.</p>
- </div>
- <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
- <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/" alt="ibd_figure" class="shadowed-image" style="width: 90%; max-width: 800px;">
- <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 3 ttrR</p>
+ <h3>Adhesion Protein Als3</h3>
+ <p>Agglutinin-like sequence protein3 (Als3) is a cell surface glycoprotein of Candida albicans that plays a crucial role in vitro adhesion and biofilm formation. Als3 is essential for the binding of Candida albicans hyphae to various host cell surface receptor proteins, and it induces endocytosis by binding to E-cadherin on epithelial cells (Wang Tianming et al., Candida-epithelial interactions, 2023). Since our chassis organism, Saccharomyces cerevisiae, is also a fungus like Candida albicans, expressing Als3 on the surface of Saccharomyces cerevisiae cells is expected to enable binding to E-cadherin on the small intestinal epithelium, thereby allowing the engineered bacteria to colonize the small intestine.</p>
+ <p>To verify the effectiveness of Als3, we designed an adhesion assay experiment. We designed the corresponding plasmids to express Als3 in Saccharomyces cerevisiae and simultaneously used the expression of EGFP for yeast tracking. After transforming the plasmids into Saccharomyces cerevisiae, we selected yeast that had been successfully transformed using Ura nutrient-deficient medium. Then, we verified the effectiveness of Als3 through the adhesion capability assay. For further details, please refer to the protocol</p>
+ <p>Aim:</p>
+ <p>To verify the change in adhesion capability of Saccharomyces cerevisiae expressing Als3 to small intestinal cells </p>
+ <p>Constructs:</p>
+ <p>pYES2-SV40-ALS3-EGFP</p>
+ <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
+ <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/pic6.png" alt="ibd_figure" class="shadowed-image" style="width: 40%; max-width: 400px;">
+ <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 6 pYES2-SV40-ALS3-EGFP plasmid</p>
+ </div>
+ <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
+ <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/pic7.jpg" alt="ibd_figure" class="shadowed-image" style="width: 40%; max-width: 400px;">
+ <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 7 the result of transformation</p>
+ </div>
+ <p>In our colonization system, we designed to express TtrSR and Als3 in Saccharomyces cerevisiae. Als3 is located downstream of the ttrB promoter. After expressing this system in Saccharomyces cerevisiae, when tetrathionate is present in the intestine (i.e., at the site of IBD lesions), the tetrathionate sensor is activated, leading to the expression of Als3 protein. This ultimately allows the engineered bacteria to adhere to the intestinal epithelial cells, thereby achieving colonization. </p>
+ <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
+ <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/pic8.png" alt="ibd_figure" class="shadowed-image" style="width: 40%; max-width: 400px;">
+ <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/pic9.png" alt="ibd_figure" class="shadowed-image" style="width: 40%; max-width: 400px;">
+ <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 8,9 Final construction of the colonization system</p>
+ </div>
</div>
- <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
- <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/ alt="ibd_figure" class="shadowed-image" style="width: 90%; max-width: 800px;">
- <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 4 ttrS</p>
- </div>
- <p>To test the effectiveness of Als3, we designed a bacterial attachment experiment. We constructed the corresponding plasmid and after transforming it into Saccharomyces cerevisiae, we used human intestinal sections to test the adhesion effects and differences between S. cerevisiae expressing Als3 and yeast without Als3 expression. For more details, please refer to the protocol.</p>
- <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
- <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/" alt="ibd_figure" class="shadowed-image" style="width: 90%; max-width: 800px;">
- <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 5 Als3</p>
- </div>
- <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
- <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/ alt="ibd_figure" class="shadowed-image" style="width: 90%; max-width: 800px;">
- <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 6 转化åŽ</p>
- </div>
</div>
<div class="row mt-4">
@@ -117,46 +135,35 @@
<h2 id="Result">
<h2>Result</h2>
<hr>
- </div>
- </div>
-
- <div class="row mt-4">
- <div class="col-lg-12">
+ <p>Through our efforts, we have obtained some experimental results to support our system design. We divided the colonization system into the TtrRS system and the Als3 protein and tested their functionality through a series of wet lab experiments. Finally, we plan to integrate them to test the effectiveness of the complete colonization system. You can see our experimental results in the following content.</p>
<h3>Tetrathionate sensor TtrSR</h3>
- <p>After acquiring the fluorescence confocal microscopy images, we calculated the ratio of total fluorescence intensity to the number of cells.</p>
- </div>
- <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
- <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/" alt="ibd_figure" class="shadowed-image" style="width: 90%; max-width: 800px;">
- <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 7 Confocal images of one group with induction and one without</p>
- </div>
- <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
- <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/" alt="ibd_figure" class="shadowed-image" style="width: 90%; max-width: 800px;">
- <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 8 Statistical graph</p>
+ <p>After successfully transforming the pYES2-SV40-ttrS and pESC-SV40-ttrR-PttrB-EGFP plasmids into Saccharomyces cerevisiae, we selected three colonies with good conditions for amplification. Then, the same clone was divided into two groups, one group was induced with 1mM tetrathionate, and the other group was used as a control without inducer. After 12 hours of induction, an appropriate amount of bacterial solution was prepared into temporary slides and the fluorescence intensity was tested using a fluorescence confocal microscope. After obtaining the fluorescence confocal microscopy images, we calculated the ratio of total fluorescence intensity to the number of cells. The images were processed using Image J, and statistical graphs were plotted using GraphPad Prism.</p>
+ <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
+ <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/pic10.png" alt="ibd_figure" class="shadowed-image" style="width: 60%; max-width: 600px;">
+ <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 10 Fluorescence confocal microscopy imaging results (s: control group without inducer; s+: K<sub>2</sub>O<sub>6</sub>S<sub>4</sub> added)</p>
+ </div>
+ <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
+ <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/pic11.png" alt="ibd_figure" class="shadowed-image" style="width: 60%; max-width: 600px;">
+ <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 11 Statistical results of tetrathionate induction experiment (s: control group without inducer; s+: K<sub>2</sub>O<sub>6</sub>S<sub>4</sub> added)</p>
+ </div>
+ <p>The results showed that after tetrathionate induction, the expression of the downstream protein EGFP increased, but there was no significant difference compared to the control group. Meanwhile, in the S+ group, although EGFP was expressed, the fluorescence was weak, which means the expression level was low</p>
+ <p>To explore the reasons for the insignificant induction difference and the low expression level, we conducted further literature research and group discussions. We finally concluded that this might be due to compatibility issues that arise when the tetrathionate sensor TtrSR is transplanted from E.coli to Saccharomyces cerevisiae. Due to differences in protein expression and delivery systems between eukaryotic and prokaryotic cells, TtrS may not effectively function in Saccharomyces cerevisiae due to issues with signal peptides, transmembrane domains, etc., which may prevent it from properly localizing to the cell membrane. Additionally, the expression levels of TtrS and TtrR in Saccharomyces cerevisiae may be insufficient. During the literature research, we found that some researches have achieved the application of two-component systems in eukaryotic systems through codon optimization. In following experiments, we plan to optimize the TtrS protein, including codon optimization, signal peptide prediction and optimization, membrane localization prediction, and transmembrane domain optimization, in order to enable TtrRS to function better in Saccharomyces cerevisiae.</p>
+ <h3>Adhesion Protein Als3</h3>
+ <p>We used human small intestinal sections to test the adhesion effects and differences between Saccharomyces cerevisiae expressing Als3 and yeast without Als3 expression. The yeast expressing Als3 was transformed with the pYES2-SV40-ALS3-EGFP plasmid, while the yeast without Als3 expression was also transformed with plasmid that could express EGFP, allowing for the observation of Saccharomyces cerevisiae. The human small intestinal sections were purchased from Zhongke Guanghua Company. After the adhesion assay, we stained the sections with Hoechst 33342. This staining allowed the small intestinal cells to emit blue fluorescence. Therefore, we could observe the small intestinal tissue through the blue fluorescence, while the green fluorescence marked the position of Saccharomyces cerevisiae. We counted the yeast cells adhered to the tissue and calculated the ratio of cell number to tissue area. The images were processed using Image J, and statistical graphs were plotted using GraphPad Prism.</p>
+ <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
+ <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/pic12.png" alt="ibd_figure" class="shadowed-image" style="width: 60%; max-width: 600px;">
+ <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 12 <b>luorescence confocal microscopy imaging results of attachment experiment</b>. a, saccharomyces cerevisiae expressing Als3. b, wild- type saccharomyces cerevisiae (without Als3, control group). c, intestinal tissue section with Als3-expressing saccharomyces cerevisiae adhesion. d, intestinal tissue section with wild-type saccharomyces cerevisiae adhesion</p>
+ </div>
+ <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
+ <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/pic13.png" alt="ibd_figure" class="shadowed-image" style="width: 60%; max-width: 600px;">
+ <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 13 <b>Statistical results of quantification of attachment experiment</b> (Als3: the attachment of saccharomyces cerevisiae expressing Als3; WT: the attachment of wild-type saccharomyces cerevisiae, control groups)</p>
+ </div>
+ <p>The results showed that after expressing Als3, there was a significant difference in the number of Saccharomyces cerevisiae adhering per unit area of small intestinal epithelium compared to the wild-type group, with an increase in adhesion. This indicates that Als3 can enhance the adhesion ability of Saccharomyces cerevisiae to small intestinal tissue to some extent. However, during the observation of the images, we found that the green fluorescence of Saccharomyces cerevisiae was not stable, with some yeast showing weak or even no fluorescence. Additionally, there was spontaneous fluorescence within the small intestinal tissue, which could affect the counting of yeast cells. </p>
+ <p>Therefore, in subsequent experiments, we plan to use cell dyes such as Trypan Blue for more uniform and accurate staining. Moreover, to better test the adhesion effect of the engineered bacteria, we plan to conduct further in vivo experiments using mice. Furthermore, since it seems that expressing Als3 does not significantly increase the number of adherent cells, we also plan to compare or optimize using other adhesion proteins or systems.</p>
</div>
- <p>The results indicate that after induction, the expression of the downstream protein increased, but there was no significant difference compared to the control group as anticipated.</p>
- <p>This could be due to compatibility issues that arise when the tetrathionate sensor TtrSR is transferred from E. coli to Saccharomyces cerevisiae. Because of differences between eukaryotic and prokaryotic protein expression and delivery systems, TtrS may not be properly processed and localized to the cell membrane in S. cerevisiae. In subsequent experiments, we optimized the TtrS protein, including codon optimization, structural modeling and prediction (still being modified——, to be added later).</p>
</div>
- <div class="row mt-4">
- <div class="col-lg-12">
- <h3>Adhesive protein Als3</h3>
- <p>After conducting the attachment experiment, we counted the yeast cells left on the tissue and calculated the ratio of the number of cells to the tissue area.</p>
- </div>
- <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
- <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/" alt="ibd_figure" class="shadowed-image" style="width: 90%; max-width: 800px;">
- <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 9 Tissue staining image</p>
- </div>
- <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
- <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/" alt="ibd_figure" class="shadowed-image" style="width: 90%; max-width: 800px;">
- <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 8 Light microscopy image</p>
- </div>
- <div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
- <img src="https://static.igem.wiki/teams/5187/wiki-colonization-fig/" alt="ibd_figure" class="shadowed-image" style="width: 90%; max-width: 800px;">
- <p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 10 Statistical graph</p>
- </div>
- <p>The results indicate that after expressing Als3, the adhesion ability of Saccharomyces cerevisiae to intestinal tissue has significantly improved. It has been demonstrated that Als3 can indeed be used for colonization in Saccharomyces cerevisiae.</p>
- </div>
<div class="row mt-4">
<div class="col-lg-12">
@@ -197,6 +204,9 @@
<div class="col-lg-12">
<h2 id="References">
<h2>References</h2>
+ <p>[1] Levitt MD, Furne J, Springfield J, Suarez F, DeMaster E. Detoxification of hydrogen sulfide and methanethiol in the cecal mucosa. J Clin Invest. 1999 Oct;104(8):1107-14. doi: 10.1172/JCI7712. PMID: 10525049; PMCID: PMC408582.</p>
+ <p>[2] Daeffler KN, Galley JD, Sheth RU, Ortiz-Velez LC, Bibb CO, Shroyer NF, Britton RA, Tabor JJ. Engineering bacterial thiosulfate and tetrathionate sensors for detecting gut inflammation. Mol Syst Biol. 2017 Apr 3;13(4):923. doi: 10.15252/msb.20167416. PMID: 28373240; PMCID: PMC5408782.</p>
+ <p>[3] Wächtler B, Citiulo F, Jablonowski N, Förster S, Dalle F, Schaller M, Wilson D, Hube B. Candida albicans-epithelial interactions: dissecting the roles of active penetration, induced endocytosis and host factors on the infection process. PLoS One. 2012;7(5):e36952. doi: 10.1371/journal.pone.0036952. Epub 2012 May 14. PMID: 22606314; PMCID: PMC3351431.</p>
<hr>
</div>
</div>
--
GitLab