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Information × Registration Number 0219U102065, 0119U103151 , R & D reports Title Structural and functional characteristics of Scots pine defensins as candidate genes for control of crop diseases. popup.stage_title Head Kovaleva Valentina A., Кандидат біологічних наук Registration Date 17-12-2019 Organization Ukrainian national forestry university popup.description2  We developed the protocol for the expression of defensin2 from Scots pine in the pET32 / BL21-CodonPlus (DE3) -RIL expression system and obtaining the target soluble peptide conjugated with thioredoxin (TRX). It was established that the highest yield of soluble TRX-PsDef2 is achieved by the induction of transformed cells with 0.5 mM isopropyl-β-D-1-thiogalactopyranoside (IPTG) and culturing them for 3 hours at 25 ° C. Chimeric TRX-PsDef2 protein was purified on HisPurNi-NTA resin and TRX-tag was subsequently removed by proteolytic cleavage with enterokinase. PsDef2 was isolated from the proteolytic mixture by ion-exchange chromatography on an SP-Sepharose Fast Flow column using a step-gradient from 0.05 to 1 M NaCl. We obtained one milligram of the target peptide with purity higher than 95% from one liter of bacterial culture. It was found that recombinant peptide PsDef2 has activity against phytopathogenic fungus Fusariumsporotrichiellaand oomycetesPhythophtoragonapodyides at concentrations 5.0μM. Product Description popup.authors popup.nrat_date 2020-04-02 Close