Genetic analysis in Salmonella typhimurium with a small collection of randomly spaced insertions of transposon Tn10 delta 16 delta 17

Abstract

We report the isolation of a group of 279 Salmonella typhimurium strains carrying randomly spaced insertions of the minitransposon Tn10 delta 16 delta 17 and describe the use of these strains to facilitate genetic analysis. The insertions were isolated initially in individual recombinant lambda clones from a genomic library. Individual insertions were then moved into the S. typhimurium chromosome, where the distribution of insertion sites relative to standard genetic markers was analyzed in a series of transductional crosses. Since a different, randomly chosen clone was used to generate each insertion, the distribution of insertion positions should have been as random as the cloning events leading to the formation of the library. In agreement with this expectation, most S. typhimurium markers tested were cotransducible with one or more of these Tn10 delta 16 delta 17 insertions. We expect that most new mutations will be quickly classified and mapped by determination of the pattern of cotransduction with this set of insertions. This use is illustrated by the analysis of a group of lac operon fusions regulated by anaerobiosis. We also describe several other applications that should make this collection a useful new tool in S. typhimurium genetics.