A xylose-inducible Bacillus subtilis integration vector and its application

Gene
L KimW Schumann

Abstract

The construction of a xylose-inducible expression vector is described. This vector allows the integration of any gene, coding for its authentic protein, at the amyE locus of Bacillus subtilis (Bs). The controlable expression cassette consists of the repressor-encoding gene and the promoter of the Bacillus megaterium-derived operon for xylose utilization, sandwiched between the 5'- and 3'-ends of amyE. This thereby allows insertion of in vitro constructed transcriptional fusions at the amyE locus of the Bs chromosome. The versatility of this expression system was tested by fusing three different heat-shock genes to the xylose-inducible promoter and following their expression by Western immunoblot analysis. Whereas no increase in the amount of heat-shock protein could be detected under non-inducing conditions when compared to the isogenic wild-type strain, the three proteins were strongly induced after addition of xylose, depending on the gene. To determine the tightness and the induction factor of the system more accurately, the bgaB gene encoding a heat-stable beta-galactosidase (beta Gal) was analyzed. The background activity of beta Gal increased by a factor of at least 200 after addition of xylose. The system is not subject ...Continue Reading

References

Jul 1, 1992·Biochimie·C Karmazyn-CampelliP Stragier
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Oct 15, 1958·Proceedings of the National Academy of Sciences of the United States of America·J Spizizen

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Citations

Jul 13, 2007·Current Microbiology·Hoang Duc NguyenWolfgang Schumann
Oct 18, 2000·FEMS Microbiology Letters·T M WendrichM A Marahiel
Nov 14, 1997·Current Opinion in Biotechnology·W M de VosO P Kuipers
Nov 6, 2010·The Journal of Biological Chemistry·Marcus MiethkeMohamed A Marahiel
Sep 10, 2009·Antimicrobial Agents and Chemotherapy·Thijs R H M KouwenJan Maarten van Dijl
Dec 3, 2008·Antimicrobial Agents and Chemotherapy·Jean-Yves F DuboisJan Maarten van Dijl
Nov 8, 2006·Applied and Environmental Microbiology·Elise DarmonJan Maarten van Dijl
Jun 6, 2003·Applied and Environmental Microbiology·David A SchofieldMichael G Schmidt
Dec 13, 2006·Infection and Immunity·Patrick H Tu QuocWilliam L Kelley
May 24, 2005·Infection and Immunity·Bing Wang, Howard K Kuramitsu
Apr 17, 2007·Journal of Bacteriology·Adrian J JervisAnne Moir
Jun 22, 2006·Journal of Bacteriology·Marcus MiethkeUlf Gerth
Aug 4, 2009·Journal of Bacteriology·Falk HillmannHubert Bahl
Jan 26, 2010·Journal of Bacteriology·Alexander G AlbrechtMohamed A Marahiel
Mar 3, 2009·Journal of Bacteriology·Diana García-MorenoMontserrat Elías-Arnanz
Apr 28, 2001·Journal of Bacteriology·R MeimaM E Lidstrom
Aug 10, 2002·Journal of Bacteriology·Isabelle GuillouardIsabelle Martin-Verstraete
Dec 18, 2003·Journal of Bacteriology·Ulf GerthMichael Hecker
Jun 19, 2004·Journal of Bacteriology·Annette KamionkaWolfgang Hillen
Aug 20, 2005·Journal of Bacteriology·Urszula Zielenkiewicz, Piotr Ceglowski
Oct 22, 2011·Microbial Cell Factories·Kambiz Morabbi HeraviJosef Altenbuchner

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