@@ -179,55 +179,91 @@ The engineering success since when we compare the result is more significant whe
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<h2>Testing TurboID-AIP toxicity to human cells</h2>
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<b>Purpose:</b><br><br>
The purpose of the experiment is to evaluate the damage of <i>S. aureus</i> enterotoxin toward human cells, and the amount of enterotoxin produced by Biotinylated <i>S. aureus</i>. We intended to use the apoptosis rate of epithelial cell to reflect the production of enterotoxin.
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<b>Design:</b><br><br>
We divided <i>S. aureus</i> into two groups. One is the control group with lysate extracted only from <i>S. aureus</i>. Another is a lysate extracted from TurboID-AIP and biotin blocked S.aureus.
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<b>Materials:</b><br><br>
1. Inner lining epithelial cell sample in human mouth<br><br>
2. Culture media containing bovine serum<br><br>
3. <i>S. aureus</i> DNA <br><br>
4. Biotin powder<br><br>
5. <i>S. aureus</i> Lysate<br><br>
6. Biotin labeled S aureus Lysate<br><br>
<b>S. aureus Lysate preparation:</b><br><br>
1. Extract 1ml of <i>S. aureus</i> (OD0.3)<br><br>
2. <i>S. aureus</i> in bovine serum lysed via supersonic schizolysis method.<br><br>
4. Collect 1ml of <i>S. aureus</i> (OD0.3) after centrifugation.<br><br>
5. <i>S. aureus</i> in bovine serum lysed via supersonic schizolysis method.<br><br>
<b>Fluid preparation for flow cytometry:</b><br><br>
1. Scrape epithelial cells with a dentiscalprum.<br><br>
2. Separate the sample into two parts.<br><br>
3. Mix the samples with <i>S. aureus</i> Lysate and Biotin labeled <i>S. aureus</i> lysate respectively (10 minutes).<br><br>
4. Centrifugation.<br><br>
5. Discard the supernatant.<br><br>
6. Resuspend the cell pellet for 15 minutes.<br><br>
7. Collect precipitated cells (1400rpm) after centrifugation.<br><br>
8. Analyze the fluid with flow cytometry. <br><br>
<b>Result:</b><br><br>
It is presented that in figure 1, 3, 5, the enterotoxin damages the epithelial cells. In figure 2, 4, 6, the apoptosis rate of cells declined indicating that the damage of enterotoxin decreased. Thus, we can conclude that our project can successfully block the production of enterotoxin in <i>S. aureus</i>.<br><br>
Fig.1 Analysis of epithelial cell of subject 1 with <i>S. aureus</i> lysate from (A) to (C); (A) size of the cells. (B) width of cells. It eliminates the duplicates which are useless to the Analysis. (C) The apoptosis rate of cells.
Fig. 5 测试者2 上皮细胞和添加TurboID-AIP, biotin工程菌及 <i>S. aureus</i>超声裂解液混合10 min
Fig.2 Analysis of epithelial cell of subject 1 with biotin labeled <i>S. aureus</i> lysate from (A) to (C); (A) size of the cells. (B) width of cells. It eliminates the duplicates which are useless to the Analysis. (C) The apoptosis rate of cells.
Fig.3 Analysis of epithelial cell of subject 2 with <i>S. aureus</i> lysate from (A) to (C); (A) size of the cells. (B) width of cells. It eliminates the duplicates which are useless to the Analysis. (C) The apoptosis rate of cells.
Fig. 7 测试者3 上皮细胞和添加TurboID-AIP, biotin工程菌及 <i>S. aureus</i>超声裂解液混合10 min
Fig.4 Analysis of epithelial cell of subject 2 with biotin labeled <i>S. aureus</i> lysate from (A) to (C); (A) size of the cells. (B) width of cells. It eliminates the duplicates which are useless to the Analysis. (C) The apoptosis rate of cells.
Fig.5 Analysis of epithelial cell of subject 3 with <i>S. aureus</i> lysate from (A) to (C); (A) size of the cells. (B) width of cells. It eliminates the duplicates which are useless to the Analysis. (C) The apoptosis rate of cells.
Fig.6 Analysis of epithelial cell of subject 3 with biotin labeled <i>S. aureus</i> lysate from (A) to (C); (A) size of the cells. (B) width of cells. It eliminates the duplicates which are useless to the Analysis. (C) The apoptosis rate of cells
