I need methods to check to see if a plasmid that I conjugated into a bacterial species has been altered such that the function of the plasmid is knocked out. I'm looking for methods beyond doing just a test digest.
Bacterial Conjugation
Conjugation is merely the fusion of two compatible bacterial cells. Bringing two genotypes together and allowing them to conjugate is the equivalent of making a cross in eukaryotes. Our discussion of conjugation will center on the gut bacterium Escherichia coli (E. coli). Conjugation and gene transfer in E. coli are driven by a circular DNAplasmid called the fertility factor or sex factor (F), which is found in some but not all cells. Hence to understand how to make a cross in E. coli, we have to understand the properties of F.
Check the full of version
https://www.ncbi.nlm.nih.gov/books/NBK21351/
My issue is not with conjugation. I have already demonstrated successful conjugation delivery of my plasmid. The issue is that some colonies are escaping the effects of my plasmid, and I need to know what they're doing to it in order to achieve that.
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