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Hi. I'm trying to integrate heterologous genes into E.coli genome.
The problem is that the size of the genes are about 7 kb.
The only system that I can use is a home-made suicide vector using R6K origin and sacB.
Several trials ended up with failures.
Do you think that 7 kb is too long to integrate into E.coli genome with this system?
Actually, I don't have any experiences to mutate E.coli. Please give me any tips of advice. (incubation time, cautions, troubleshootings etc.)
Thank you in advance.
