PMID: 9163905May 1, 1997Paper

Integrative vector for constructing single-copy translational fusions between regulatory regions of Bacillus subtilis and the bgaB reporter gene encoding a heat-stable beta-galactosidase

FEMS Microbiology Letters
O StossWolfgang Schumann

Abstract

Here we report on the construction of two integrative plasmids for Bacillus subtilis allowing in vitro construction of translational fusions. Both plasmids contain two cassettes in tandem: the bgaB gene encoding a heat-stable beta-galactosidase devoid of its own regulatory sequences and the first two codons followed by a neomycin-resistance gene for selection in B. subtilis. Both cassettes are flanked by the 3'- and 5'-end of the amyE gene (encoding alpha-amylase) allowing integration of both cassettes at the amyE locus of the B. subtilis chromosome. For propagation in Escherichia coli, the plasmids contain the pBR322 origin of DNA replication and the beta-lactamase-encoding gene. Whereas one vector needs a promoter, a Shine-Dalgarno sequence and the beginning of a gene fused in-frame to bgaB, the other one already carries a constitutive promoter. The versatility of the gene fusion vectors was demonstrated by the integration of the regulatory regions of the dnaK and the cat-86 genes. In the first case, heat-inducible expression was found, and by comparison with an operon fusion, it seems that the dnaK operon is regulated at both the transcriptional and the posttranscriptional level. In the second case, chloramphenicol-inducible...Continue Reading

References

Jun 1, 1986·Journal of Bacteriology·H HirataH Okada
Feb 1, 1996·Molecular Microbiology·M HeckerU Völker

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Citations

Jul 25, 2007·Molecular Genetics and Genomics : MGG·Torsten WaldminghausFranz Narberhaus
Jun 26, 1999·Molecular Microbiology·G HomuthW Schumann
Jun 20, 2007·Nucleic Acids Research·Nadja HeidrichSabine Brantl
Jul 17, 2007·Molecular Microbiology·Torsten WaldminghausFranz Narberhaus

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