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Current Microbiology

, Volume 52, Issue 2, pp 143–148 | Cite as

Versatile Expression and Secretion Vectors for Bacillus subtilis

  • Ulf Brockmeier
  • Marion Wendorff
  • Thorsten EggertEmail author
Article

Abstract

Most expression systems are based on Escherichia coli as the host strain because of the large availability of all kinds of vector plasmids. However, aside from the obvious advantages of E. coli systems, serious problems can occur during the process of heterologous gene expression and purification. Therefore, low expression rates, formation of inclusion bodies, improper protein-folding, and/or toxicity problems might enforce changing the expression host. Here we describe the construction of two new vectors, pBSMuL1 and pBSMuL2, for overexpression and secretion of heterologous proteins in Bacillus subtilis as an alternative expression host. The new plasmids combine several advantages in comparison to available Bacillus expression systems: an appropriate multiple cloning site consisting of 13 unique restriction sites, one (pBSMuL1) or two (pBSMuL2) strong constitutive promoters, a high efficient signal sequence for protein secretion, and the possibility to express proteins as His-tagged fusions for easy detection and purification. We have demonstrated the applicability of the novel vector plasmids for the production and purification of the heterologous cutinase from Fusarium solani pisi.

Keywords

Fusarium Solani Efficient Secretion Heterologous Gene Expression Strong Constitutive Promoter Secretion Vector 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

One author (U. B.) was supported by a scholarship from the European Graduate College 795 entitled Regulatory Circuits in Cellular Systems: Fundamentals and Biotechnological Applications, which was funded by the Deutsche Forschungsgemeinschaft.

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Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Ulf Brockmeier
    • 1
  • Marion Wendorff
    • 1
  • Thorsten Eggert
    • 1
    Email author
  1. 1.Institut für Molekulare EnzymtechnologieHeinrich-Heine Universität DüsseldorfForschungszentrum JülichGermany

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