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| <h4>Module 3- Activates in presence of salicylate</h4> | | <h4>Module 3- Activates in presence of salicylate</h4> |
| </section><br /> | | </section><br /> |
− | <center><div style="img-data"><img src="https://static.igem.org/mediawiki/2017/9/99/T--IISER-Mohali-INDIA--circuit2.png" alt="Figure1. Genetic circuit for detection of salicylate" width="100%" height="100%"></div></center> | + | <center><div class="data-img"><img src="https://static.igem.org/mediawiki/2017/9/99/T--IISER-Mohali-INDIA--circuit2.png" alt="Figure1. Genetic circuit for detection of salicylate" width="100%" height="100%"></div></center> |
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| <section><h3>To clone three modules in the <i>E.coli</i>, they should have different copy number plasmids. The module 1 will be active even in the absence of pollutant. So, we decided to put it on low copy number plasmid. The module 2 will get activated only after pollutant is present. So, we decided to put it on high copy number plasmid for immediate detection. The module 3 is already present in the <i>E.coli</i> chromosome.</h3><br/> | | <section><h3>To clone three modules in the <i>E.coli</i>, they should have different copy number plasmids. The module 1 will be active even in the absence of pollutant. So, we decided to put it on low copy number plasmid. The module 2 will get activated only after pollutant is present. So, we decided to put it on high copy number plasmid for immediate detection. The module 3 is already present in the <i>E.coli</i> chromosome.</h3><br/> |
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| </section><br /> | | </section><br /> |
| <section><h3>Both the constructs were cloned in low copy number plasmid, pZS21MCS. In the presence of T7 RNA polymerase, chromoprotein II is transcribed and translated into bacteria giving it yellow color. Construct 1 is cloned at AatII and ApaI site, and construct 2 is cloned at SalI and BamHI site.</h3></section> | | <section><h3>Both the constructs were cloned in low copy number plasmid, pZS21MCS. In the presence of T7 RNA polymerase, chromoprotein II is transcribed and translated into bacteria giving it yellow color. Construct 1 is cloned at AatII and ApaI site, and construct 2 is cloned at SalI and BamHI site.</h3></section> |
− | <center><div class="img-data"><img src="https://static.igem.org/mediawiki/2017/a/a5/T--IISER-Mohali-INDIA--GD2.png" alt="Figure 2: Vector map of pZS21MCS with cloned module 1" | + | <center><div class="data-img"><img src="https://static.igem.org/mediawiki/2017/a/a3/T--IISER-Mohali-INDIA--GDsa.png" alt="Figure 2: Vector map of pZS21MCS with cloned module 1" |
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| <section> | | <section> |
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| </section> | | </section> |
| <section><h3>Both constructs were cloned in intermediate copy plasmid, pACYC177. Construct 1 was cloned at XhoI and XmaI site and construct 2 was cloned at BamHI and AatII site.</h3></section><br /> | | <section><h3>Both constructs were cloned in intermediate copy plasmid, pACYC177. Construct 1 was cloned at XhoI and XmaI site and construct 2 was cloned at BamHI and AatII site.</h3></section><br /> |
− | <center><<div class="img-data"> | + | <center><div class="data-img"> |
− | <img src="https://static.igem.org/mediawiki/2017/e/e2/T--IISER-Mohali-INDIA--GD3.png" alt="Figure 3: Vector map of pACYC177 with cloned module 2" width="100%" height="100%" ></div></center> <br/> | + | <img src="https://static.igem.org/mediawiki/2017/6/6e/T--IISER-Mohali-INDIA--GDsb.png" alt="Figure 3: Vector map of pACYC177 with cloned module 2" width="100%" height="100%" ></div></center> <br/> |
| <section><h3>Module 3 :</h3> | | <section><h3>Module 3 :</h3> |
| <h4>Module 3 contains <i>mar</i> promoter which is native to <i>E. coli</i> strain and present in mar operon. MarRAB operon is present in <i>E. coli<\i>. It has three genes - <i>marR</i>, <i>marA</i> and <i>marB</i>. <i>marR</i> promoter is present upstream of marR and has two pair of direct repeat elements nearby.</h4> | | <h4>Module 3 contains <i>mar</i> promoter which is native to <i>E. coli</i> strain and present in mar operon. MarRAB operon is present in <i>E. coli<\i>. It has three genes - <i>marR</i>, <i>marA</i> and <i>marB</i>. <i>marR</i> promoter is present upstream of marR and has two pair of direct repeat elements nearby.</h4> |