Proposed Construction of pBAD24-OmpA for the Differential Expression of OmpA in Conjugation Efficiency Studies

A better understanding of the mechanisms behind bacterial conjugation may allow for selectively enhancing or repressing conjugation; this has possible profound implications, both clinically and industrially. OmpA, an abundant outer membrane protein of Escherichia coli, has been shown to be necessary for conjugation and mating-pair formation, although the details of this interaction have not been fully elucidated. Although an o mpA gene containing pCR2.1- TOPO plasmid has been developed, a system allowing for the controlled and differential expression of OmpA would be more appropriate for investigating the effect of increasing OmpA levels on conjugation efficiency. The current investigation attempted to clone ompA into a pBAD24 vector in order to produce a pBAD24-OmpA construct. Although ompA was amplified and the plasmid was isolated, no transformants were observed as pBAD28 was accidentally received in place of pBAD24. Despite this error, the reported investigation describes appropriate protocols for the correct construction of the pBAD24-OmpA vector system.