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Team:Oxford/Results
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| - | + | Wetlab Results: <br> | |
| + | For the bioremediation aspect of DCMation, we have managed to:<br> | ||
| + | 1. purify the pUNI-ABTUNJK and pSB1C3-ABTUNJK plasmids, which encode the microcompartments<br> | ||
| + | 2. verify expression of pUNI-ABTUNJK plasmids in E. coli using Western blotting<br> | ||
| + | 3. insert the ABTUNJK vector into pSRKGm using Gibson assembly, for expression in P. putida <br> | ||
| + | 4. insert the dcmA gene into the pCM66 backbone using Gibson assembly, since pCM66 is suitable for expression in Methylobacterium extorquens DM4 <br> | ||
| + | 5. insert dcmA into the pRSFDuet, and transformed this into DH5α cells for hypermutagenic PCR on the dcmA gene <br> | ||
| + | 6. fuse dcmA and sfGFP using PCR, followed by restriction and ligation to insert the dcmA-sfGFP transcriptional fusion into pRSFDuet. This was then transformed into DH5α cells and imaged using fluorescence microscopy <br> | ||
| + | 7. insert microcompartment-tagged dcmA into pRSFDuet<br> | ||
| + | 8. insert the microcompartment-tagged sfGFP into pME6010, and used fluorescence microscopy to image this<br> | ||
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Revision as of 15:13, 16 October 2014
