Mutant isolation and cloning of the gene encoding protease VII from Escherichia coli

Abstract

A mutant of Escherichia coli lacking protease VII, the outer membrane-associated protease which specifically cleaves paired basic residues (1), was isolated by using N-methyl-N'-nitro-N-nitrosoguanidine treatment. The mutant exhibited no significant change as for its growth rate and microscopic feature compared with wild cells. The gene encoding protease VII was cloned by using complementation analysis of protease VII (-) mutation. The minicell experiment showed that the gene encoded a putative precursor protein of 38,000 Mr which was processed into a protein of 36,000 Mr suggesting the presence of a signal peptide on the putative precursor.