Difference between revisions of "Team:JiangnanU China/Design"

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<h1>Design</h1>
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Design is the first step in the design-build-test cycle in engineering and synthetic biology. Use this page to describe the process that you used in the design of your parts. You should clearly explain the engineering principles used to design your project.
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<h3>What should this page contain?</h3>
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            src: url('https://static.igem.org/mediawiki/2019/4/49/T--JiangnanU_China--FuturaFuturisC-Italic.otf');
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<li>Explanation of the engineering principles your team used in your design</li>
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        }
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<li>Discussion of the design iterations your team went through</li>
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<li>Experimental plan to test your designs</li>
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</ul>
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<h3>Inspiration</h3>
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<ul>
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<li><a href="https://2016.igem.org/Team:MIT/Experiments/Promoters">2016 MIT</a></li>
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<li><a href="https://2016.igem.org/Team:BostonU/Proof">2016 BostonU</a></li>
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<li><a href="https://2016.igem.org/Team:NCTU_Formosa/Design">2016 NCTU Formosa</a></li>
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                <div class="fb_72">
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                    <b>Design</b>
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                </div>
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                            <div class="fb_48" style="margin-left: 2%;margin-top: 1%">View all</div>
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                Recombinant <i>E. coli</i> resistant to phage infection will be constructed. It will be mainly divided into four
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                parts.
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                1. Looking for Inducible Promoters
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            <div class="fm_22">
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                Therefore, transcriptome data from different stages of phage infection will be measured to find parts that
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                can respond to phage infection at latent period and burst period. Fluorescence gene <i>gfp</i> and
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                <i>mCherry</i>
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                will be used to characterize them.
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            <img src="https://static.igem.org/mediawiki/2019/d/db/T--JiangnanU_China--design3.png"
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                2. Looking for Anti-phage Protein
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            <div class="fm_22">
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                On the one hand, we will search for resistant parts that can resist phage infection through literature, and
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                use plate test to determine the resistance effect of the parts.
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                <br/><br/>
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                On the other hand, we will use ARTP (Atmospheric and Room Temperature Plasma) mutagenesis screening to screen
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                for bacteriophage-resistant parts. Specifically, we identify the mutant strain by co-culture with the
 +
                phage, and after sorting out the mutant strain, we culture all the mutant strains for ten generations
 +
                to strengthen the mutant sites. In this process, the phage plate test has been carried out to eliminate
 +
                the degraded resistant strains.
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            </div>
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            <img src="https://static.igem.org/mediawiki/2019/f/f3/T--JiangnanU_China--project_designs_2.png"
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                Finally, we will obtain some mutant strains and we will select key sites by comparing the whole genome.
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            <img src="https://static.igem.org/mediawiki/2019/3/3b/T--JiangnanU_China--design4.png"
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                Anti-phage detection is carried out on the selected anti-phage part, and the part with the best
 +
                anti-phage effect is cascaded with the anti-phage part screened in the literature, and both of them
 +
                are connected to the inducible promoter that could respond to phages in the latent period.
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                3. Kill Switch
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            <br/>
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            <div class="fm_22">
 +
                In the second part, we plan to find anti-phage parts which could in the latent period resist to phage.
 +
                However, if the phage skip our first line of defense, we are able to ligate the kill switch with the
 +
                burst period inducible promoter to kill the cell before the complete assembly of phage.
 +
            </div>
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            <div class="split_small"></div>
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            <img src="https://static.igem.org/mediawiki/2019/c/cc/T--JiangnanU_China--project_designs_4.png"
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                4. Application
 +
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 +
            <br/>
 +
            <div class="fm_22">
 +
                When the recombinant mutant is constructed, we will revisit our original purpose, which is to play a role in the practical application. In view of the advantages of our school Jiangnan University in fermentation engineering, we will apply our construction circuit to the production strain to verify its ability to resist phage. This experiment will be done in a specific laboratory where our school works with the respective companies, and we will ensure the safety of the entire experiment and prevent any bacteria and phage from leaking.
 +
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{{:Team:JiangnanU_China/Footer}}

Latest revision as of 19:01, 21 October 2019

JiangNan

Recombinant E. coli resistant to phage infection will be constructed. It will be mainly divided into four parts.
1. Looking for Inducible Promoters

Therefore, transcriptome data from different stages of phage infection will be measured to find parts that can respond to phage infection at latent period and burst period. Fluorescence gene gfp and mCherry will be used to characterize them.
2. Looking for Anti-phage Protein

On the one hand, we will search for resistant parts that can resist phage infection through literature, and use plate test to determine the resistance effect of the parts.

On the other hand, we will use ARTP (Atmospheric and Room Temperature Plasma) mutagenesis screening to screen for bacteriophage-resistant parts. Specifically, we identify the mutant strain by co-culture with the phage, and after sorting out the mutant strain, we culture all the mutant strains for ten generations to strengthen the mutant sites. In this process, the phage plate test has been carried out to eliminate the degraded resistant strains.
Finally, we will obtain some mutant strains and we will select key sites by comparing the whole genome.
Anti-phage detection is carried out on the selected anti-phage part, and the part with the best anti-phage effect is cascaded with the anti-phage part screened in the literature, and both of them are connected to the inducible promoter that could respond to phages in the latent period.
3. Kill Switch

In the second part, we plan to find anti-phage parts which could in the latent period resist to phage. However, if the phage skip our first line of defense, we are able to ligate the kill switch with the burst period inducible promoter to kill the cell before the complete assembly of phage.
4. Application

When the recombinant mutant is constructed, we will revisit our original purpose, which is to play a role in the practical application. In view of the advantages of our school Jiangnan University in fermentation engineering, we will apply our construction circuit to the production strain to verify its ability to resist phage. This experiment will be done in a specific laboratory where our school works with the respective companies, and we will ensure the safety of the entire experiment and prevent any bacteria and phage from leaking.
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