Development of techniques for the genetic manipulation of the gliding bacteriaLysobacter enzymogenesandLysobacter brunescens

Abstract

Lysobacter enzymogenes and Lysobacter brunescens are Gram-negative gliding bacteria that belong to the γ subgroup of the proteobacteria. As a first step toward a molecular analysis of Lysobacter gliding motility, we developed techniques to genetically manipulate these bacteria. Cosmid pSUP106 of the broad host range incompatibility group Q (Inc Q) was introduced into L. enzymogenes and L. brunescens by conjugation and electroporation. pSUP106 replicated stably in both organisms and conferred antibiotic resistance. We also identified several other plasmids (pKT210, pH1JI) that functioned in L. enzymogenes and a transposon (mini-Tn5Sp) that functioned in L. brunescens. The identification of these tools allows genetic analysis of Lysobacter gliding motility, exoenzyme production, and production of antibiotics and other secondary metabolites.Key words: Lysobacter, gliding motility, gene transfer.