We've started working with Acetobacters.
A commercial broad-range shuttle vector looked propagated in this bacterium;
- confirmed by 1)colony formation on antibiotic-containing agars, 2) retransformation test of isolated plasmids into E.coli and 3) multiple copy-number (qRT-PCR)
(I cound not see a single band from isolated plasmid samples, because of all the smear)
However, one colleague doubted on this shuttle vector.
He said that when a native gene was cloned in this shuttle vector, it was incorporated into the genome like a suicide vector via single cross-over. That is, this shuttle vector does not replicate in the bacterium.
However, he can not explain the cases that no native gene was cloned in the shuttle vector and transformed successfully.
Is it possible that one shuttle vector propagated by its origin of replication and at the same time, it functions as a suicide vector under some conditions?
The replication of origin may not be strong or stable.
Thank you for your help in advance.