TU Darmstadt / ULB / TUbiblio

Production of recombinant and tagged proteins in the hyperthermophilic archaeon Sulfolobus solfataricus.

Albers, Sonja-Verena and Jonuscheit, Melanie and Dinkelaker, S. and Urich, Tim and Kletzin, Arnulf and Tampé, R. and Driessen, A. J. M. and Schleper, Christa (2006):
Production of recombinant and tagged proteins in the hyperthermophilic archaeon Sulfolobus solfataricus.
72, In: Applied and environmental microbiology, (1), pp. 102-11, ISSN 0099-2240, [Article]

Abstract

Many systems are available for the production of recombinant proteins in bacterial and eukaryotic model organisms, which allow us to study proteins in their native hosts and to identify protein-protein interaction partners. In contrast, only a few transformation systems have been developed for archaea, and no system for high-level gene expression existed for hyperthermophilic organisms. Recently, a virus-based shuttle vector with a reporter gene was developed for the crenarchaeote Sulfolobus solfataricus, a model organism of hyperthermophilic archaea that grows optimally at 80 degrees C (M. Jonuscheit, E. Martusewitsch, K. M. Stedman, and C. Schleper, Mol. Microbiol. 48:1241-1252, 2003). Here we have refined this system for high-level gene expression in S. solfataricus with the help of two different promoters, the heat-inducible promoter of the major chaperonin, thermophilic factor 55, and the arabinose-inducible promoter of the arabinose-binding protein AraS. Functional expression of heterologous and homologous genes was demonstrated, including production of the cytoplasmic sulfur oxygenase reductase from Acidianus ambivalens, an Fe-S protein of the ABC class from S. solfataricus, and two membrane-associated ATPases potentially involved in the secretion of proteins. Single-step purification of the proteins was obtained via fused His or Strep tags. To our knowledge, these are the first examples of the application of an expression vector system to produce large amounts of recombinant and also tagged proteins in a hyperthermophilic archaeon.

Item Type: Article
Erschienen: 2006
Creators: Albers, Sonja-Verena and Jonuscheit, Melanie and Dinkelaker, S. and Urich, Tim and Kletzin, Arnulf and Tampé, R. and Driessen, A. J. M. and Schleper, Christa
Title: Production of recombinant and tagged proteins in the hyperthermophilic archaeon Sulfolobus solfataricus.
Language: English
Abstract:

Many systems are available for the production of recombinant proteins in bacterial and eukaryotic model organisms, which allow us to study proteins in their native hosts and to identify protein-protein interaction partners. In contrast, only a few transformation systems have been developed for archaea, and no system for high-level gene expression existed for hyperthermophilic organisms. Recently, a virus-based shuttle vector with a reporter gene was developed for the crenarchaeote Sulfolobus solfataricus, a model organism of hyperthermophilic archaea that grows optimally at 80 degrees C (M. Jonuscheit, E. Martusewitsch, K. M. Stedman, and C. Schleper, Mol. Microbiol. 48:1241-1252, 2003). Here we have refined this system for high-level gene expression in S. solfataricus with the help of two different promoters, the heat-inducible promoter of the major chaperonin, thermophilic factor 55, and the arabinose-inducible promoter of the arabinose-binding protein AraS. Functional expression of heterologous and homologous genes was demonstrated, including production of the cytoplasmic sulfur oxygenase reductase from Acidianus ambivalens, an Fe-S protein of the ABC class from S. solfataricus, and two membrane-associated ATPases potentially involved in the secretion of proteins. Single-step purification of the proteins was obtained via fused His or Strep tags. To our knowledge, these are the first examples of the application of an expression vector system to produce large amounts of recombinant and also tagged proteins in a hyperthermophilic archaeon.

Journal or Publication Title: Applied and environmental microbiology
Volume: 72
Number: 1
Divisions: 10 Department of Biology > Sulfur Biochemistry and Microbial Bioenergetics
?? fb10_mikrobiologie ??
10 Department of Biology
Date Deposited: 24 May 2011 08:18
Export:
Suche nach Titel in: TUfind oder in Google
Send an inquiry Send an inquiry

Options (only for editors)

View Item View Item