Biopolym. Cell. 2008; 24(5):377-384.
Structure and Function of Biopolymers
Production of Scots pine recombinant defensin 1 and its antifungal activity
1Kovalyova V. A., 1Gout R. T., 2Gout I. T.
  1. National Forestry University of Ukraine
    103, General Chuprynky, Lviv, Ukraine, 79057
  2. University College London
    Gower Str., London WC1E 6BT, UK


Recently we have purified an endogenous defensin from Scots pine germs, and cloned cDNA encoding defensin 1 (PsDef1, Pinus sylvestris defensin 1). The cDNA region encoding a mature form of Scots pine defensin 1 was cloned into a vector pET 42a(+), and the expression of recombinant GST/PsDef1 in the Escherichia coli bacterial system was induced. The conditions of production of soluble GST-proteins were optimized. After purification of the recombinant protein by affinity chromatography on Glutathione-Sepharose column and proteolytic cleavage with Factor Xa, the functionally active preparation of recombinant PsDef1 was obtained. Its antifungal activity is similar to that of endogenous Scots pine defensin 1.
Keywords: recombinant defensin PsDef1, expression, affinity purification, antifungal activity


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