Difference between revisions of "Team:ECUST/Biocide"
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<img src="https://static.igem.org/mediawiki/2018/b/b2/T--ECUST--BiocideF1.png" | <img src="https://static.igem.org/mediawiki/2018/b/b2/T--ECUST--BiocideF1.png" | ||
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<p>We insert fragment of Cecropin AD into vector pET-28a(Figure 1) </p> | <p>We insert fragment of Cecropin AD into vector pET-28a(Figure 1) </p> | ||
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<img src="https://static.igem.org/mediawiki/2018/9/9e/T--ECUST--result--biocide_1.jpg" class="f1"> | <img src="https://static.igem.org/mediawiki/2018/9/9e/T--ECUST--result--biocide_1.jpg" class="f1"> | ||
<figcaption><b>Figure 1. The vector pET-28a Vector is cut by NcoI and BamHI. Sequence of AD was chemically synthesized and amplified by PCR, then ligated with linearized vector by Ezmax.</b></figcaption> | <figcaption><b>Figure 1. The vector pET-28a Vector is cut by NcoI and BamHI. Sequence of AD was chemically synthesized and amplified by PCR, then ligated with linearized vector by Ezmax.</b></figcaption> | ||
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<p>The plasmid was transformed to E. coli BL21 and cultured at 37 °C for 12 h. positive monoclonal bacteria were cultured and verified by PCR(Figure 2). </p> | <p>The plasmid was transformed to E. coli BL21 and cultured at 37 °C for 12 h. positive monoclonal bacteria were cultured and verified by PCR(Figure 2). </p> | ||
<figure> | <figure> | ||
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<img src="https://static.igem.org/mediawiki/2018/4/47/T--ECUST--result--biocide_2.jpg" class="f1"> | <img src="https://static.igem.org/mediawiki/2018/4/47/T--ECUST--result--biocide_2.jpg" class="f1"> | ||
<figcaption><b>Figure 2. 1% Agarose Gel Electrophoresis of PCR, which shows that our vector was successfully constructed.</b></figcaption> | <figcaption><b>Figure 2. 1% Agarose Gel Electrophoresis of PCR, which shows that our vector was successfully constructed.</b></figcaption> | ||
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<p>We verified the expression of Cecropin AD by SDS-PAGE(Figure 3). </p> | <p>We verified the expression of Cecropin AD by SDS-PAGE(Figure 3). </p> | ||
<figure> | <figure> | ||
| − | <figure class="makeresponsive" style="width: 50%;"> | + | <figure class="makeresponsive" style="width: 50%;margin-left:23%;margin-right:30%;"> |
<img src="https://static.igem.org/mediawiki/2018/4/46/T--ECUST--result--biocide_3.jpg" class="f3"> | <img src="https://static.igem.org/mediawiki/2018/4/46/T--ECUST--result--biocide_3.jpg" class="f3"> | ||
<figcaption><b>Figure 3. The SDS-PAGE of Cecropin AD | <figcaption><b>Figure 3. The SDS-PAGE of Cecropin AD | ||
Revision as of 22:14, 17 October 2018
Description
When the concentration of Fe3+ in the environment is high, it means that the hard skeleton of the microenvironment of the iron bacteria is destroyed and loses protection. At this time, the added bactericide will have a greater killing effect on the bacteria. The bactericide we selected was cecropin AD, a cationic chimeric antimicrobial peptide obtained from cecropin, consisting of the first 11 amino acid residues of cecropin A and the last 26 amino acid residues of cecropin D. It has strong antibacterial activity against Gram-positive and negative bacteria.
Design
Result
We insert fragment of Cecropin AD into vector pET-28a(Figure 1)
The plasmid was transformed to E. coli BL21 and cultured at 37 °C for 12 h. positive monoclonal bacteria were cultured and verified by PCR(Figure 2).
We verified the expression of Cecropin AD by SDS-PAGE(Figure 3).
