Difference between revisions of "Team:NRP-UEA-Norwich/Results"
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<p>Attempted a triparental conjugation and additional electroporation of plasmids containing <i>HydABC</i> Strep-II tag on C-terminus and <i>HydABC</i> Strep-II tag on N-terminus constructs into wild type, double knock out and FeFe knock out <i>Shewanella oneidensis</i> MR-1</p> | <p>Attempted a triparental conjugation and additional electroporation of plasmids containing <i>HydABC</i> Strep-II tag on C-terminus and <i>HydABC</i> Strep-II tag on N-terminus constructs into wild type, double knock out and FeFe knock out <i>Shewanella oneidensis</i> MR-1</p> | ||
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Revision as of 02:07, 19 October 2016
Results
Cloning Results
Cloning the FeFe and NiFe hydrogenase genes individually into iGEM BioBricks, which can then be cloned into pBAD vector using Golden Gate Cloning (GGC).
Protein Modelling
Modelling hydrogenases and MtrCAB in Shewanella oneidensis using homology sequencing.
BioBrick Conjugation into Shewanella oneidensis
In order to assess whether our BioBricks would function in our chassis organism Shewanella oneidensis MR-1 we attempted to transform and express two BioBricks into S. oneidensis MR-1; J04450 containing a gene for the red fluorescent protein (RFP) and K584001 which contains a gene for the green fluorescent protein (GFP). These BioBricks were chosen as they contain reporter genes and successfully transformed colonies could be easily identified under UV light.
Triparental conjugation and electroporation of plasmids into Shewanella oneidensis
Attempted a triparental conjugation and additional electroporation of plasmids containing HydABC Strep-II tag on C-terminus and HydABC Strep-II tag on N-terminus constructs into wild type, double knock out and FeFe knock out Shewanella oneidensis MR-1
Measuring the Optical Density of the Shewanella Oneidensis knockout strain (LS473) and Wildtype strain (MR-1) with a media control
Date: 18th - 19th August 2016
Aim: The aim of the experiment is to measure the optical density of both the MR-1 shewanella oneidensis strains (MR1 and LS473) when stored anaerobically and quantify the amount of hydrogen in the headspace gas using gas chromatography.
Sponsors
