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Applied Microbiology and Biotechnology

, Volume 33, Issue 6, pp 657–663 | Cite as

Regulated expression of heterologous genes inBacillus subtilis using the Tn10 encodedtet regulatory elements

  • Manfred Geissendörfer
  • Wolfgang Hillen
Applied Genetics and Regulation

Summary

TheEscherichia coli-derivedtet regulatory elements from Tn10 have been used to construct vectors allowing the regulated, inducible, high-level expression of foreign genes inBacillus subtilis. While the wild-typetet promoters are inactive inB. subtilis, a synthetic mutanttet sequence with improved promoter consensus sequences and upstream poly A blocks shows activity inB. subtilis. The expression of an indicatorcat gene is inducible by sublethal amounts of tetracycline, indicating that the Tet repressor protein and thetet operator sequences are functional. However, the inducibility and maximal expression are not sufficient in this construct. To improve these properties atet operator sequence was placed between the —35 and —10 boxes of theB. subtilis-derived very strongxyl promoter. In the presence of atetR gene this construct is about 100-fold inducible and has high promoter strength, but some basal expression. This is avoided by placing a secondtet operator downstream resulting in no detectable basal expression at the expense of reduced inducibility. Using the system with a singletet operator inducible expression of glucose dehydrogenase fromB. megaterium was obtained at a very high level, and inducible expression of human single-chain urokinase-like plasminogen activator was achieved at the same level as inE. coli. Unlike inE. coli, the product was not degraded up to 4 h after induction inB. subtilis. These results demonstrate that the regulated expression vector described here should be very useful for production of foreign gene products fromB. subtilis cultures.

Keywords

Plasminogen Regulatory Element Foreign Gene Basal Expression Inducible Expression 
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.

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Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • Manfred Geissendörfer
    • 1
  • Wolfgang Hillen
    • 1
  1. 1.Lehrstuhl für Mikrobiologie, Institut für Mikrobiologie und BiochemieFriedrich-Alexander Universität Erlangen-NürnbergErlangenFederal Republic of Germany

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