Cloning of the structural gene (ompA) for an integral outer membrane protein of Escherichia coli K-12.

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Abstract
The gene (ompA) for the major outer membrane protein II* from E. coli K-12 was cloned on a 5-megadalton EcoRI fragment, using phage .lambda. as vector. The gene is expressed during the lytic cycle of the recombinant phage and the insoluble membrane-bound protein was detected in phage plaques with a simple radioimmunoassay. Transfer of the EcoRI fragment into plasmid pSC101 and expression in a host lacking protein II* led to overproduction of protein II* and decreased production of 2 other major outer membrane proteins. Expression of the plasmid pSC101-ompA+ in minicells derived from an ompA minicell-producing strain led to synthesis, at high rates, of this protein and massive accumulation of a 2nd cell envelope protein most likely representing the biosynthetic precursor of protein II*.