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Plasmid pGEX-5T: An alternative system for expression and purification of recombinant proteins

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Summary

A novel expression vector pGEX-5T was constructed which directs the synthesis of a fusion protein with a histidine-hexapeptide and glutathione-S-transferase at its N-terminus and the recombinant protein at its C-terminus inEscherichia coli. The designed fusion gene strategy allows the purification of soluble and insoluble recombinant proteins to homogeneity with single-step affinity chromatography using immobilized glutathione and metal chelating matrix, respectively. The principle and availability of this new expression system was respectively tested with the purification of a soluble and insoluble recombinant fusion protein containing 24 and 75 amino acids of the human thrombomodulin.

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Authors and Affiliations

  1. Department of Molecular Biology, ELIAS Entwicklungslabor, D-7800, Freiburg, Germany

    Heike Berthold, Brigitte Frorath, Mirko Scanarini, Charles C. Abney & Wolfgang Northemann

  2. Institute of Clinical Chemistry and Laboratory Diagnosis, University of Rostock, O-2500, Rostock, Germany

    Bruno Ernst

Authors
  1. Heike Berthold
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  2. Brigitte Frorath
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  3. Mirko Scanarini
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  4. Charles C. Abney
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  5. Bruno Ernst
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  6. Wolfgang Northemann
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Berthold, H., Frorath, B., Scanarini, M. et al. Plasmid pGEX-5T: An alternative system for expression and purification of recombinant proteins. Biotechnol Lett 14, 245–250 (1992). https://doi.org/10.1007/BF01022318

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  • DOI: https://doi.org/10.1007/BF01022318

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