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

 
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                     <b>Design</b>
 
                     <b>Design</b>
 
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                    <div class="split"></div>
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                    <!--                View more-->
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                    <a href="#phage" style="text-decoration: none">
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                        <div class="row" style="align-content: center;color: white;">
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                            <img
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                                    src="https://static.igem.org/mediawiki/2019/0/09/T--JiangnanU_China--host_liubianxing.png"
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                            <div class="fb_48" style="margin-left: 2%;margin-top: 1%">View all</div>
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             <div class="split_small"></div>
 
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             <img src="https://static.igem.org/mediawiki/2019/8/89/T--JiangnanU_China--project_designs_0.png"
 
             <img src="https://static.igem.org/mediawiki/2019/8/89/T--JiangnanU_China--project_designs_0.png"
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            <img src="https://static.igem.org/mediawiki/2019/3/3f/T--JiangnanU_China--RESEARCH_IDEA.png"
 
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             <div class="split_small"></div>
 
             <div class="split_small"></div>
 
             <div class="fm_22">
 
             <div class="fm_22">
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                 Recombinant <i>E. coli</i> resistant to phage infection was constructed. It is mainly divided into four
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                 Recombinant <i>E. coli</i> resistant to phage infection will be constructed. It will be mainly divided into four
 
                 parts.
 
                 parts.
 
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                 First Part-looking for Inducible Promoters
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                 1. Looking for Inducible Promoters
 
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             <div class="fm_22">
 
             <div class="fm_22">
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                 Therefore, transcriptome data from different stages of phage infection were measured to find parts that
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                 Therefore, transcriptome data from different stages of phage infection will be measured to find parts that
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                 could respond to phage infection at latent period and burst period Fluorescence gene <i>gfp</i> and
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                 can respond to phage infection at latent period and burst period. Fluorescence gene <i>gfp</i> and
 
                 <i>mCherry</i>
 
                 <i>mCherry</i>
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                 were used to characterize them.
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                 will be used to characterize them.
 
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             <img src="https://static.igem.org/mediawiki/2019/0/04/T--JiangnanU_China--project_designs_1.png"
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             <img src="https://static.igem.org/mediawiki/2019/d/db/T--JiangnanU_China--design3.png"
 
                 style="width: 100%;height: auto;">
 
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                 Second Part-looking for Anti-phage Protein
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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 searched for resistant parts that can resist phage infection through literature, and
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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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                 used plate test to determine the resistance effect of the parts.
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                 use plate test to determine the resistance effect of the parts.
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                 On the other hand, we use ARTP (Atmospheric and Room Temperature Plasma) mutagenesis screening to screen
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                 for bacteriophage-resistant parts. Specifically, we identified the mutant strain by co-culture with the
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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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                 phage, and after sorting out the mutant strain, we cultured all the mutant strains for ten generations
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                 for bacteriophage-resistant parts. Specifically, we identify the mutant strain by co-culture with the
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                 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
 
                 to strengthen the mutant sites. In this process, the phage plate test has been carried out to eliminate
 
                 the degraded resistant strains.
 
                 the degraded resistant strains.
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             <div class="split_small"></div>
 
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             <div class="fm_22">
 
             <div class="fm_22">
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                 Finally, four phage-resistant mutant strains were obtained. By comparing the whole genome, we selected
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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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                key sites.
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             <img src="https://static.igem.org/mediawiki/2019/2/27/T--JiangnanU_China--project_designs_3.png"
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             <img src="https://static.igem.org/mediawiki/2019/3/3b/T--JiangnanU_China--design4.png"
 
                 style="width: 100%;height: auto;">
 
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             <div class="split_small"></div>
 
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             <div class="fm_22">
 
             <div class="fm_22">
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                 Anti-phage detection was carried out on the selected anti-phage part, and the part with the best
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                 Anti-phage detection is carried out on the selected anti-phage part, and the part with the best
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                 anti-phage effect was cascaded with the anti-phage part screened in the literature, and both of them
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                 anti-phage effect is cascaded with the anti-phage part screened in the literature, and both of them
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                 were connected to the inducible promoter that could respond to phages in the latent period.
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                 are connected to the inducible promoter that could respond to phages in the latent period.
 
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                 Third Part-kill Switch
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                 3. Kill Switch
 
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             <div class="fm_22">
 
             <div class="fm_22">
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                 In the second part, we were to find anti-phage parts which could in the latent period resist to phage.
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                 In the second part, we plan to find anti-phage parts which could in the latent period resist to phage.
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                 However, if the phage skip our first line of defense, we were able to ligated the kill switch to the
+
                 However, if the phage skip our first line of defense, we are able to ligate the kill switch with the
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                 burst period inducible promoter, killing the cell before the complete assembly of phage.
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                 burst period inducible promoter to kill the cell before the complete assembly of phage.
 
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                 Fourth Part-application
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                 4. Application
 
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             <div class="fm_22">
 
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                 The constructed recombinant <i>E. coli</i> is applied to produce γ-aminobutyric acid in 5 L and 30 L
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                 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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                fermentation cultures in the laboratory.
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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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