Integrative vectors for constructing single-copy transcriptional fusions between Bacillus subtilis promoters and various reporter genes encoding heat-stable enzymes

Gene
A MogkW Schumann

Abstract

Here, we report on the construction of three integrative plasmids for Bacillus subtilis (Bs) allowing in vitro construction of transcriptional fusions. These plasmids contain a neomycin- or tetracycline-resistance cassette and one of three promoterless genes: bgaB (encoding beta-galactosidase), cat (chloramphenicol acetyltransferase), or xylE (catechol 2,3-dioxygenase). All cassettes are flanked by the 3'- and 5'-ends of the amyE gene (encoding alpha-amylase) allowing integration of these cassettes at the amyE locus of the Bs chromosome. For propagation and selection in Escherichia coli, the plasmids contain the pBR322 origin of DNA replication and the beta-lactamase-encoding bla gene. Four unique restriction sites can be used for insertion of restriction fragments carrying promoter fragments. All three reporter genes express heat-stable enzymes (stable up to at least 50 degrees C for 30 min) as shown here. We would like to point to the modular nature of these plasmids where the three reporter genes and the two resistance cassettes can be combined in any permutation. The versatility of the promoter-probe vectors was demonstrated by the integration of the promoters of the dnaK and groE operons of Bs and following their heat-indu...Continue Reading

Citations

Nov 5, 2011·Applied Microbiology and Biotechnology·Norma WelschThomas Schweder
Jul 29, 2009·Current Microbiology·Trang Thi Phuong Phan, Wolfgang Schumann
Jun 12, 2003·FEMS Microbiology Letters·Thomas Wiegert, Wolfgang Schumann
Feb 6, 2004·FEMS Microbiology Letters·Matthias KotschwarWolfgang Schumann
Apr 19, 2012·Molecular BioSystems·Ulf W LiebalOlaf Wolkenhauer
May 16, 2002·Journal of Applied Microbiology·E MellyP Setlow
Jun 26, 1999·Molecular Microbiology·G HomuthW Schumann
Jul 17, 2001·Molecular Microbiology·T WiegertW Schumann
Jun 8, 2001·Journal of Bacteriology·T Wiegert, W Schumann
Aug 5, 2003·Journal of Bacteriology·Akram Atalla, Wolfgang Schumann
Jan 4, 2003·Journal of Bacteriology·Saskia VersteegWolfgang Schumann
Jul 28, 2013·Microbial Cell Factories·Johannes KabischThomas Schweder
Aug 30, 2005·Protein Expression and Purification·Trang Thi Phuong PhanWolfgang Schumann
Aug 11, 2006·Molecular Microbiology·Stephan ZellmeierThomas Wiegert
Apr 9, 2015·Applied Microbiology and Biotechnology·Norma WelschThomas Schweder
Feb 20, 2007·Protein Expression and Purification·Ai Thi Thuy Le, Wolfgang Schumann
Jul 14, 2012·Journal of Biotechnology·Johannes KabischThomas Schweder
Feb 5, 2005·Anais Da Academia Brasileira De Ciências·Luís C S FerreiraWolfgang Schumann
Sep 28, 1999·Journal of Bacteriology·G HomuthD Jahn
Jun 6, 1998·Journal of Bacteriology·A MogkU Völker
Feb 1, 1997·Journal of Bacteriology·G HomuthW Schumann

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