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Application of TEV Protease in Protein Production

  • Deborah A. Polayes
  • T. Dawn Parks
  • Stephen A. Johnston
  • William G. Dougherty
Part of the Methods in Molecular Medicine™ book series (MIMM, volume 13)

Abstract

In many cases, the analysis of a specific protein is impeded by the inability to purify large amounts of it from a native source. Proteins of interest may be present in minute quantities and/or purification may be plagued with technical problems. Recombinant DNA methodologies have enabled researchers to circumvent some of these limitations by producing and purifying large quantities of protein in a nonnative system. Various systems and strategies have been successfully employed, depending on the specific protein of interest and the desired use of the final end product (antibody production, crystallography studies etc.). This chapter reviews some common methods for the production of recombinant fusion proteins and specifically describes a versatrle method for the removal of affinity tags from recombinant fusions using a highly purified proteinase with an unparalleled degree of specificity. This proteinase, from the genome of tobacco etch virus (TEV), demonstrates specific proteolytic activity under a wide range of parameters (salt, temperature, pH), making it an excellent choice for cleavage of fusion proteins (1,2).

Keywords

Fusion Protein Tobacco Etch Virus Protease Cleavage Site Tobacco Etch Virus Protease Chemical Cleavage 
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

© Humana Press Inc. 1998

Authors and Affiliations

  • Deborah A. Polayes
    • 1
  • T. Dawn Parks
    • 2
  • Stephen A. Johnston
    • 3
  • William G. Dougherty
    • 2
  1. 1.Molecular Biology Research and DevelopmentLife Technologies, Inc.Gaithersburg
  2. 2.Department of MicrobiologyOregon State UniversityCorvallis
  3. 3.Department of BiochemistryUniversity of Texas Southwestern Medical CenterDallas

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