Suicide plasmid vehicles for insertion mutagenesis in Rhizobium meliloti and related bacteria

Abstract

The construction and use of a set of plasmid vectors of the transposons Tn1, Tn5 and Tn9 that are suicidal in Rhizobium spp. and suitable for mutagenesis with these 3 transposons are described. The vectors are composed of the p15A replicon which functions in Escherichia coli but not in Rhizobium spp. and a region encoding the N type of bacterial conjugation system which is very efficient in matings between E. coli and Rhizobium spp. The usefulness of the vectors was most extensively assessed in R. meliloti. It is likely that they will be useful for mutagenesis and genome manipulation in other bacteria as well.