"
Team:Dundee/Results
From 2012.igem.org
(Difference between revisions)
Nici Morrice (Talk | contribs) |
Nici Morrice (Talk | contribs) |
||
| Line 71: | Line 71: | ||
<center><h2><img src="https://static.igem.org/mediawiki/2012/2/20/Lab.jpg"></h2></center><br> | <center><h2><img src="https://static.igem.org/mediawiki/2012/2/20/Lab.jpg"></h2></center><br> | ||
| - | <u>Characterisation<u > | + | <u>Characterisation<u /> |
The thirteen gene components that make up the type VI secretion system (T6SS) were successfully cloned into two separate PT7.5 pUNI-PROM vectors and are as follows:<br> | The thirteen gene components that make up the type VI secretion system (T6SS) were successfully cloned into two separate PT7.5 pUNI-PROM vectors and are as follows:<br> | ||
The overall aim of this project is that of targeting C. difficile for removal through the attachment of an endolysin specific towards C. difficile onto the T6SS the team had cloned. Not only was this endolysin specifically attached onto the tip of the T6SS (VgrG) but it was also possible to fuse the endolysin gene onto the shaft (Hcp) of the T6SS too. Upon characterisation of these fusions, bands were not visible. It was then assumed that expression of these genes was not substantial enough to be visible and thus a T7 promoter was used and induced through the addition of IPTG and then bands for these fusions were clearly seen on SDS-PAGE gels and western blots.<br> | The overall aim of this project is that of targeting C. difficile for removal through the attachment of an endolysin specific towards C. difficile onto the T6SS the team had cloned. Not only was this endolysin specifically attached onto the tip of the T6SS (VgrG) but it was also possible to fuse the endolysin gene onto the shaft (Hcp) of the T6SS too. Upon characterisation of these fusions, bands were not visible. It was then assumed that expression of these genes was not substantial enough to be visible and thus a T7 promoter was used and induced through the addition of IPTG and then bands for these fusions were clearly seen on SDS-PAGE gels and western blots.<br> | ||
Revision as of 10:27, 25 September 2012
Characterisation The thirteen gene components that make up the type VI secretion system (T6SS) were successfully cloned into two separate PT7.5 pUNI-PROM vectors and are as follows:
The overall aim of this project is that of targeting C. difficile for removal through the attachment of an endolysin specific towards C. difficile onto the T6SS the team had cloned. Not only was this endolysin specifically attached onto the tip of the T6SS (VgrG) but it was also possible to fuse the endolysin gene onto the shaft (Hcp) of the T6SS too. Upon characterisation of these fusions, bands were not visible. It was then assumed that expression of these genes was not substantial enough to be visible and thus a T7 promoter was used and induced through the addition of IPTG and then bands for these fusions were clearly seen on SDS-PAGE gels and western blots.
