Team:Austin UTexas/Results
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.
