Integrative elements for Bacillus subtilis yielding tetracycline-dependent growth phenotypes.

ABSTRACT: We describe the construction and application of elements for random insertion of promoter containing DNA into the genome of Bacillus subtilis. The outward-facing promoter of these integrative elements termed InsTet(G+) is inducible by tetracycline so that conditional mutants are generated. We constructed three InsTet(G+) variants using different regulatory windows. In the first, the regulator gene tetR is located within the element, allowing one-step mutagenesis. The second contains tetR in the chromosome and yields the best regulation efficiency. The third exploits xylose-dependent tetR expression from a plasmid, enabling induction of TetR synthesis so that distinct expression levels of an affected gene can be adjusted. We have obtained mutant strains with all three variants. For some of

SUBMITTER: Bertram R 

PROVIDER: S-EPMC1253839 | biostudies-literature | 2005 Oct

REPOSITORIES: biostudies-literature