Checked plates that had tripartite conjugation. A number of the plate had colony growth on them!
Each of these plates were incubated at 37C on MYPGP Poly60Kan50 for three days, however growth was only observed on plates that were conjugated previously at 37C. Below is a table illustrating the amount of plates that had a number of colonies on them after the three day incubation.
| Temp | Disc | Broth |
| RT | 0/3 | 0/3 |
| 30C | 0/3 | 0/3 |
| 37C | 3/3 | 3/3 |
Below are images of three of the plates that had a number of colonies on them.
Discussion
It would appear that the tripartite conjugation was a success. That we are able to conjugate the transposon of pMarA into P. larvae 9545 using the pRK2013 helper plasmid. I will determine if the present colonies are indeed P. larvae using the diagnostic AFB primers and I would also like to determine if the original pMarA plasmid is no longer present in its entirety and that its transposon has been inserted into the P. larvae genome.
//EWW
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