Difference between revisions of "Team:Westminster/Results"
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| − | Our | + | Our initial aim was to express the MtrCAB operon found in <i>Shewanella oneidensis</i> MR-1 in <i>Escherichia coli</i>. Another objective was to investigate the potential benefits of the proteins CymA and OmcA within the microbial fuel cell. From reviewing the literature we quickly discovered that cloning these genes into <i>E.coli</i> would be a challenge as they can be potentially toxic to the cell. Due to this we decided to try to express each gene individually with His-10 tags to show that they can be expressed in <i>E.coli</i>.<br> <br> |
| + | Another potential hurdle for our project is the fact that <i>E.coli</i> does not have the advantage of nanowires in order to transport their electrons as <i>S. oneidensis</i>. Therefore, in order to overcome this the K12 derivative, DH5α was used.<br><br> | ||
| + | Westminster team took advantage on the generous offer from IDT. In doing so, the decision was made to acquire the five genes (MtrA, MtrB, MtrC, CymA, OmcA), that were the focus of this project, synthesised as gBlocks. We were fortunate enough to be selected to use a novel cloning technique designed by, RDP (Rapid DNA Prototyping). 20 primers were ordered in order to convert the gBlocks into BioBrick and RDP formats. These were designed in SnapGene to include a GC clamp and annealing temperature of 60-70°C. <br><br> | ||
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<img src=" https://static.igem.org/mediawiki/2015/0/00/Team_Westminster_Results_1.png" height="300px" width="auto"><br><br> | <img src=" https://static.igem.org/mediawiki/2015/0/00/Team_Westminster_Results_1.png" height="300px" width="auto"><br><br> | ||
Revision as of 18:48, 18 September 2015
Project Results
Our initial aim was to express the MtrCAB operon found in Shewanella oneidensis MR-1 in Escherichia coli. Another objective was to investigate the potential benefits of the proteins CymA and OmcA within the microbial fuel cell. From reviewing the literature we quickly discovered that cloning these genes into E.coli would be a challenge as they can be potentially toxic to the cell. Due to this we decided to try to express each gene individually with His-10 tags to show that they can be expressed in E.coli.
Another potential hurdle for our project is the fact that E.coli does not have the advantage of nanowires in order to transport their electrons as S. oneidensis. Therefore, in order to overcome this the K12 derivative, DH5α was used.
Westminster team took advantage on the generous offer from IDT. In doing so, the decision was made to acquire the five genes (MtrA, MtrB, MtrC, CymA, OmcA), that were the focus of this project, synthesised as gBlocks. We were fortunate enough to be selected to use a novel cloning technique designed by, RDP (Rapid DNA Prototyping). 20 primers were ordered in order to convert the gBlocks into BioBrick and RDP formats. These were designed in SnapGene to include a GC clamp and annealing temperature of 60-70°C.
After ordering these primers we ran a PCR reaction to amplify the gBlocks and add the Prefix and suffix or X and Z ends to the five genes of interest. The gels below show the results:
Click here to see details of our lab results