After getting transconjugants, I am trying to subculture but there is no growth. Out of 5 different transconjugants only 3 are growing. Is it like transconjugants loosing the stability so they are not growing? Can some please help me whats going on?
If you can provide some more details on the experiment, that would help us answer your question. Are you conjugating a plasmid, or a mobile element? Are you imposing antibiotic selection? Does the conjugated DNA have an origin of replication for the recipient?
I am working on the epidemiology of ESBL positive UTI isolates, it may be plasmid or ICE element. All I want to know is how these isolates are becoming resistant to many antibiotics? If conjugation works I can tell it is because of the mobile element.
Yes, I am imposing antibiotic selection. About ORI in the conjugated DNA, no idea.
There is no growth on control plates, transconjugants on selective antibiotic plates are not growing when I am trying to subculture.
I have previously observed this in inter-species conjugations if the mobile element cannot integrate or the plasmid does not have a suitable origin for the recipient. But this is mainly to be expected for inter-species transfer. Are your donor and recipient the same species?
Yes, both are of same species. These are clinical isolates, few are able to grow and few are not. I think it is the problem of compatibility of the ORI.
Thank you.
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