Transposon Tn5supF-based reverse genetic method for mutational analysis of Escherichia coli with DNAs cloned in λ phage

Abstract

An efficient method for systematic mutational analysis of the Escherichia coli genome was developed. It entails Tn5supF transposition to λ-E. coli hybrid phage clones (Kohara library) and then transduction of recipient cells to Sup+. Essential and nonessential genes are distinguished by the ability of insertion mutant phage to form haploid versus only heterozygous partial diploid bacterial recombinants.