title: Transposon Tn5supF-based reverse genetic method for mutational analysis of Escherichia coli with DNAs cloned in lambda phage.
abstract: An efficient method for systematic mutational analysis of the Escherichia coli genome was developed. It entails Tn5supF transposition to lambda-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.
authors.affiliation: Department of Molecular Microbiology, Washington University Medical School, St. Louis, Missouri 63110.
authors.forename: S H
authors.initials: SH
authors.lastname: Phadnis
authors.forename: S
authors.initials: S
authors.lastname: Kulakauskas
authors.forename: B R
authors.initials: BR
authors.lastname: Krishnan
authors.forename: J
authors.initials: J
authors.lastname: Hiemstra
authors.forename: D E
authors.initials: DE
authors.lastname: Berg
mesh_terms: D010582:Bacteriophage lambda / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D004252:DNA Mutational Analysis; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D010957:Plasmids; D011995:Recombination, Genetic; D012689:Sequence Homology, Nucleic Acid
chemical_list: D004251:DNA Transposable Elements; D004269:DNA, Bacterial