Team:Oxford/Results

font style="font-weight:bold">Wetlab Results:

For the font style="font-weight:bold">bioremediation aspect of DCMation, we have managed to:

1. purify the pUNI-ABTUNJK and pSB1C3-ABTUNJK plasmids, which encode the microcompartments
2. verify expression of pUNI-ABTUNJK plasmids in E. coli using Western blotting
3. insert the ABTUNJK vector into pSRKGm using Gibson assembly, for expression in P. putida
4. insert the dcmA gene into the pCM66 backbone using Gibson assembly, since pCM66 is suitable for expression in Methylobacterium extorquens DM4
5. insert dcmA into the pRSFDuet, and transformed this into DH5α cells for hypermutagenic PCR on the dcmA gene
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
7. insert microcompartment-tagged dcmA into pRSFDuet
8. insert the microcompartment-tagged sfGFP into pME6010, and used fluorescence microscopy to image this