Conjugative reporter system for the use in Bacillus licheniformis and closely related Bacilli

Abstract

Bacillus wild-type strains are genetically difficult to manipulate, and thus, the options for rational strain investigation and design are limited. Here, we present a set of small conjugative shuttle vectors for the use in Bacillus licheniformis and related, genetically difficult accessible wild-type strains. The vector set comprises the modular general-purpose vector pV2 and its derivatives pV3SDlacZ and pV3lacZ. The pV3 vectors are designed for the investigation of transcriptional and translational activities of regulatory regions like promoters and ribosomal binding sites (RBS). The vector set has been tested for investigating gene regulation under aerobic and anaerobic conditions. Significance and Impact of the StudyThe hereby introduced vectors represent a novel easy to apply toolbox which makes Bacillus strains with low or without natural competences accessible through transconjugation and allows quantitative studies on gene activity and gene regulation. Thus, the tool set enables research on newly isolated wild-type strains with methods normally limited to laboratory strains.