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[[File:T--Austin UTexas--KOM4nogfp.png|thumb|left|600px|Using a fluorescent microscope, this was a picture taken of the G. oxydans strain, KOM 4, without the plasmid that contains GFP.]] | [[File:T--Austin UTexas--KOM4nogfp.png|thumb|left|600px|Using a fluorescent microscope, this was a picture taken of the G. oxydans strain, KOM 4, without the plasmid that contains GFP.]] | ||
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+ | [[File:T--Austin UTexas--KOM4withgfpandcirclesfixed.png|thumb|left|600px|Using a fluorescent microscope, this was a picture taken of the potential transconjugant, KOM 4, with the plasmid GFP. '''16s sequencing was still needed to confirm successful conjugation.''' | ||
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Revision as of 15:49, 10 October 2016
Conjugation
We have attempted to conjugate GFP into both G. oxydans and G. hansenii with a Diaminopimelic Acid (DAP) auxotrophic strain of E. coli . The plasmid contains the vector pMMB67EH, the promoter PA-1, GFP and a spectinomycin resistance gene.
The first conjugation was done with KOM strains 4 (G. oxydans) 5 ( G. oxydans and 15 ( L. fermentati ). We attempted these conjugations before sequencing the recipient strains, so that is why we tried to conjugate into L. fermentati . First, a mixture between a KOM strain and the DAP auxotroph strain were plated on a LB+DAP solid medium to allow for conjugation to occur. After 24 hours of incubation, I scraped up the growth and plated each conjugation mixture onto a LB+Spec plate.
Next, we viewed the potential transconjugants on a fluorescence microscope.
[[File:T--Austin UTexas--KOM4withgfpandcirclesfixed.png|thumb|left|600px|Using a fluorescent microscope, this was a picture taken of the potential transconjugant, KOM 4, with the plasmid GFP. 16s sequencing was still needed to confirm successful conjugation.