Construction and characterization of F plasmid-based expression vectors

Abstract

A low-copy expression vector has been constructed from a 9 Kbp region of the Escherichia coli F plasmid containing the oriV and oriS origins of replication. This plasmid carries the beta-lactamase gene (Apr) and the araBAD promoter/araC regulator for arabinose-inducible gene expression. A derivative which carries a lacZ reporter gene was found to be stably maintained for at least 150 generations. A related multi-copy plasmid was stably maintained in arabinose-free medium, but no plasmid-bearing segregants remained after 60 generations when lacZ expression was induced. Induced expression resulted in 27% (multi-copy) and 12% (low-copy) decreases in growth rate. The uninduced levels of beta-galactosidase were 200 units (multi-copy) and 15 units (low-copy).