The genus Leptospira belongs to the order Spirochaetales and is composed of both saprophytic and pathogenic members, such as Leptospira biflexa and L. interrogans, respectively. A major factor contributing to our ignorance of spirochetal biology has been the lack of methods available for genetic analysis of these organisms. In recent years, an E. coli-L. biflexa shuttle vector has been constructed and a system for targeted mutagenesis and random transposon mutagenesis of the saprophyte L. biflexa has been developed. These studies enable the use of L. biflexa as a model bacterium among spirochetes.