Bacteriophage lambda as a delivery vector for Tn10-derived transposons in Xenorhabdus bovienii

Abstract

Xenorhabdus bovienii wild-type strains lack a functional receptor protein (LamB) in the outer membrane and as a result are unable to adsorb coliphage lambda (λ). Introduction of plasmids encoding lamB into X. bovienii T228 results in constitutive expression of LamB in the outer membrane of this organism. LamB-expressing strains of X. bovienii adsorb lambda bacteriophage particles and can be used as hosts for lambda::Tn constructs. A Tn10-derived transposon, element 9 (J. C. Way, D. Davis, D. Morisato, D. E. Roberts, and N. Kleckner, Gene 32:369-379, 1984) was used to construct a variety of insertion mutants of X. bovienii. Mutants that had altered expression of protease, lipase, DNase, dye-binding capability, and hemolytic activity, in addition to a series of auxotrophic mutants, were isolated.