Purification and Expression of DNA Segment Encoding Highly-Antigenic Polypeptide from Capsid Protein P10 of Southern Rice Black-Streaked Dwarf Virus

Abstract

    Southern rice black-streaked dwarf virus-SRBSDV is a new virus species of the genus Fijivirus in the family Reovidae which caused serious rice losses in tens of millions of hectares throughout southern China and northern and central Vietnam during 2009-2010 period. To develop sensitive and specific antibody for the diagnosis of SRBSDV based on ELISA method, a DNA fragment, namely P10.1, of the SRBSDV S10 segment, that encodes a highly-antigenic polipeptide, was cloned and expressed in E. coli. P10.1 consisting of 525 nucleotides was cloned into vector pGEM®-T, then inserted into the expression region of pET28a and transfromed into E.coli strain Rosetta to express recombinant polypeptide. Polypeptide fused with His-tag was expressed at 37°C, for 16h with 1.0 mM IPTG inducer. Using Ni-NTA chromatographic column recombinant polypeptide of about 35 kDa from cell extracts was purified. The purified polypeptide will be used as antigen for inducing specific antibody against outer capsid protein of SRBSDV.