Use of Mu phages to isolate transposon insertions juxtaposed to given genes ofEscherichia coli

Abstract

The small sizes of the DNA fragments transduced by lysates of phage Mu and of mixed lysates of Mu and mini-Mu18A-1 (an internally deleted Mu phage) provide a method for the selection of insertions of transposon Tn10 located very close to givenEscherichia coli genes. Generalized transduction with Mu lysates selected for those insertions located within 38 kilobase pairs of the gene of interest whereas insertions located within about half that distance are directly selected by use of mini-Mu phages. Use of these transduction systems avoids screening of individual colonies by phage P1 transduction for those transposon insertions closely linked to a given gene. Such insertions are most useful for localized mutagenesis and for in vitro molecular cloning.