Production of Scots pine recombinant defensin 1 and its antifungal activity

Abstract

We have recently reported affinity purification and molecular cloning of defensin (PsDef1, Pinus sylvestris defensin 1) from Scots pine germinating seeds. In this study, we describe subcloning of PsDef1 mature form into bacterial expression vector pET 42à(+) and the expression of recombinant GST/PsDef1 in Escherichia coli. The conditions for the expression and affinity purification of soluble GST/PsDef1GST were defined. Affinity purification of the recombinant GST/PsDef1 on glutathione-sepharose column and proteolytic removal of GST moiety with Factor Xa allowed us to generate functionally active preparations of recombinant PsDef1. Moreover, the antimicrobial activity of recombinant PsDef1 was found to be comparable to that of endogenous Scots pine defensin.

Expression of lectin and defensin genes in Mironovskaya 808 and Roazon wheat cultivars infected with Pseudocercosporella herpotrichoides

“…One of the classes of such compounds is plant defensins -minor peptides, micromolar concentrations [3] of which demonstrate antimicrobial activity against a wide spectrum of phytopathogenic fungi as well as agents of human diseases [4,5]. Besides, they launch signalling cascades [6] and cause morphological changes in fungal hyphae [7,8].…”