Difference between revisions of "Team:XJTLU-CHINA/Demonstrate"
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| + | <p style="font-size:20px">For the whole circuit construct, we did following experiments:</p> | ||
| + | <h2>1. Gel electrophoresis</h2> | ||
| + | </div> | ||
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| + | <img width="650px" src="https://static.igem.org/mediawiki/2017/4/43/Gel_electrophoresis.jpeg"> | ||
| + | <figcaption> | ||
| + | <strong>From left to right: Psb1c3+LL-37 (cut by PstI, uncut); psb1c3+GF-17 (cut by pstI, uncut); 1kb DNA maker; | ||
| + | pEt28a (+)+3x AMPs (uncut, cut)</strong> | ||
| + | </figcaption> | ||
| + | </figure> | ||
| + | </div> | ||
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<h2>1. Expression of LL-37 in bacteria by dot blotting</h2> | <h2>1. Expression of LL-37 in bacteria by dot blotting</h2> | ||
Revision as of 01:29, 2 November 2017
Demonstration
Aimed at examining the efficacy of the antimicrobial peptides on killing S. aureus (each experiment below were repeated 3 times).
For the whole circuit construct, we did following experiments:
1. Gel electrophoresis
1. Expression of LL-37 in bacteria by dot blotting
2. Determination of the minimal inhibitory concentration (MIC) of the three AMPs.
Both single peptide of the three AMPs and the mixture of them have conspicuous lethal effect on S. aureus.
3. Absorbance (cell density) of the biofilm of S. aureus against different concentrations of the three individual AMPs or a mixture of them
The biofilm formation of S. aureus was to a great extent inhibited by these three AMPs.
Experience and conclusions:
- Peptides has shown their antimicrobial activity on S. aureus. And they can successfully expressed in E. coli.
- AgrA, which is an essential protein in quorum sensing construct, signifies toxicity to cells during growth.
- We have trouble in obtaining enough plasmid DNA extracted from L. lactis after transformation, probably due to either low copy number or low yield from the extraction protocol.
Collaborators and Supporters
