I have been working with A. tumefaciens for several months. I have generated different vectors (with Kanamycin resistance as bacterial selection maker) that I would like to test in A. rhizogenes (strain K599). I transformed my cells by electroporation following this strain's suggested settings, but I am not getting any resistant colony. I think that the origin of replication in my vectors that works in tumefaciens might not be compatible with rhizogenes. I am using the pVS1 origin of replication. I have considered testing RK2 or pRIA4 instead. I wonder if someone from the community has worked with K599 and has any suggestions.
In the end, the people who gave me the R. rhizogenes (strain K599) did not know it was contaminated with an unknown bacteria that had taken over the original culture, so that is why the pVS1 ori was not working. Just for those who are looking for answers about the compatible oris, R. rhizogenes, and A. tumefaciens strains are compatible with the following origins of replication: RK2, pVS1, and pRIA4.
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