Transposon Tn5supF-based reverse genetic method for mutational analysis of Escherichia coli with DNAs cloned in lambda phage.
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.
Phadnis
S H
SH
Department of Molecular Microbiology, Washington University Medical School, St. Louis, Missouri 63110.
Kulakauskas
S
S
Krishnan
B R
BR
Hiemstra
J
J
Berg
D E
DE
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
D004251:DNA Transposable Elements; D004269:DNA, Bacterial