|
|
(6 intermediate revisions by 2 users not shown) |
Line 48: |
Line 48: |
| <body> | | <body> |
| <div class="container"> | | <div class="container"> |
− | <header>
| + | <a href="index.html"><img src="https://static.igem.org/mediawiki/2017/7/7b/T--IISER-Mohali-INDIA--logogeco.png" style="float: left; width:8%; padding-top: 2%;" alt="gEco"></a> |
− | <a href="index.html"><img src="https://static.igem.org/mediawiki/2017/7/7b/T--IISER-Mohali-INDIA--logogeco.png" style="float: left; width:8%; padding-top: 1%;" alt="gEco"></a>
| + | |
| | | |
− | <a href="https://www.facebook.com/igemiiserm/"><img src="https://static.igem.org/mediawiki/2017/5/5f/T--IISER-Mohali-INDIA--FB.png" style="float: right; width: 4%; margin-right: 1%; padding-top: 1%;"></a> | + | <a href="https://www.facebook.com/igemiiserm/"><img src="https://static.igem.org/mediawiki/2017/5/5f/T--IISER-Mohali-INDIA--FB.png" style="float: right; width: 4%; margin-right: 4%; padding-top: 2%; position:relative;" /></a> |
− | <a href="https://www.instagram.com/igem_iisermohali/"><img src="https://static.igem.org/mediawiki/2017/c/ce/T--IISER-Mohali-INDIA--Insta.png" style="float: right; width: 4%; margin-right: 1%; padding-top: 1%;"></a> | + | |
− | <a href="#"><img src="https://static.igem.org/mediawiki/2017/d/d7/T--IISER-Mohali-INDIA--G%2B.png" style="float: right; width: 4%; margin-right: 1%; padding-top: 1%;"></a>
| + | <a href="https://www.instagram.com/igem_iisermohali/"><img src="https://static.igem.org/mediawiki/2017/c/ce/T--IISER-Mohali-INDIA--Insta.png" style="float: right; width: 4%; margin-right: 1%; padding-top: 2%; position:relative;" /></a> |
− | <a href="https://twitter.com/iGEMIISERM"><img src="https://static.igem.org/mediawiki/2017/0/04/T--IISER-Mohali-INDIA--Tw.png" style="float: right; width: 4%; margin-right: 1%; padding-top: 1%;"></a>
| + | ' |
− | <a href="https://www.youtube.com/channel/UCXhpctjIXTHUpVCwq_ctgDg"><img src="https://static.igem.org/mediawiki/2017/3/3a/T--IISER-Mohali-INDIA--You.png" style="float: right; width: 4%; margin-right: 1%; padding-top: 1%;"></a>
| + | <a href="#"><img src="https://static.igem.org/mediawiki/2017/d/d7/T--IISER-Mohali-INDIA--G%2B.png" style="float: right; width: 4%; margin-right: 1%; padding-top: 2%; position:relative;" /></a> |
− | </header>
| + | |
| + | <a href="https://twitter.com/iGEMIISERM"><img src="https://static.igem.org/mediawiki/2017/0/04/T--IISER-Mohali-INDIA--Tw.png" style="float: right; width: 4%; margin-right: 1%; padding-top: 2%; position:relative;" /></a> |
| + | |
| + | <a href="https://www.youtube.com/channel/UCXhpctjIXTHUpVCwq_ctgDg"><img src="https://static.igem.org/mediawiki/2017/3/3a/T--IISER-Mohali-INDIA--You.png" style="float: right; width: 4%; margin-right: 1%; padding-top: 2%; position:relative;" /></a> |
| + | |
| <div class="page_logo"> | | <div class="page_logo"> |
| <img src="https://static.igem.org/mediawiki/2017/4/4d/T--IISER-Mohali-INDIA--Design.png" width="100%" height="100%"></div> | | <img src="https://static.igem.org/mediawiki/2017/4/4d/T--IISER-Mohali-INDIA--Design.png" width="100%" height="100%"></div> |
| | | |
| <article> | | <article> |
− | <div class="page-title"><h2>Genetic Design</h2> | + | <div class="page-title">Genetic Design |
| </div> <br /><br /><br /><br /><br /><br /> | | </div> <br /><br /><br /><br /><br /><br /> |
| <section><h3>The circuit for detecting the salicylate is shown in figure 1. It comprises of three modules :</h3> | | <section><h3>The circuit for detecting the salicylate is shown in figure 1. It comprises of three modules :</h3> |
Line 68: |
Line 71: |
| <h4>Module 3- Activates in presence of salicylate</h4> | | <h4>Module 3- Activates in presence of salicylate</h4> |
| </section><br /> | | </section><br /> |
− | <center><table><tr><td><img src="https://static.igem.org/mediawiki/2017/4/4a/T--IISER-Mohali-INDIA--GD1.png" alt="Figure1. Genetic circuit for detection of salicylate" width="100%" height="100%"></td></tr></table></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> |
| <br/> | | <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/> | | <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/> |
Line 77: |
Line 80: |
| </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><table><tr><td><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" |
− | width="100%" height="100%"> </td></tr></table></center> <br /> | + | width="100%" height="100%"></div></center> <br /> |
| <section> | | <section> |
| <h3>Module 2 :</h3> | | <h3>Module 2 :</h3> |
Line 86: |
Line 89: |
| </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><table><tr><td> | + | <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%" ></td></tr></table></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> |
| </section> | | </section> |
| <br/> | | <br/> |
− | <center><table><tr><td> | + | <center> |
− | <img src="https://static.igem.org/mediawiki/2017/3/35/T--IISER-Mohali-INDIA--GD4.png" alt="Figure 3: Vector map of pACYC177 with cloned module 2" width="100%" height="100%" > | + | <div class="img-data"><img src="https://static.igem.org/mediawiki/2017/3/35/T--IISER-Mohali-INDIA--GD4.png" alt="Figure 3: Vector map of pACYC177 with cloned module 2" width="100%" height="100%" ></div></center> |
− | </td></tr></table></center> <br />
| + | <br /> |
| <section><h3> | | <section><h3> |
| marR repressor is constitutively produced in <i>E. coli</i> and represses transcription of marRAB operon. Presence of salicylate inhibits marR repressor protein and induces expresssion of marRAB operon. </h3></section> | | marR repressor is constitutively produced in <i>E. coli</i> and represses transcription of marRAB operon. Presence of salicylate inhibits marR repressor protein and induces expresssion of marRAB operon. </h3></section> |