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

  Do Thi Hanh, Pham Thanh Tam, Pham Xuan Hoi

  Date Received: 26-03-2015 / Date Accepted: 09-10-2015 / Date Published: 13-06-2025

   Abstract PDF (VIETNAMESE)

  Southern rice black-streaked dwarf virus-SRBSDV isa new virus species of the genus Fijivirusin the family Reovidae which caused serious rice losses in tens ofmillions of hectares throughout southern China and northernandcentral Vietnam during2009-2010period. To developsensitive and specific antibody for the diagnosis of SRBSDV based on ELISA method, a DNAfragment, namely P10.1, of the SRBSDV S10 segment, that encodes a highly-antigenic polipeptide, was cloned and expressed in E. coli.P10.1 consistingof525 nucleotides was cloned into vector pGEM®-T, then inserted into the expression region of pET28a and transfromed into E.colistrainRosetta to express recombinant polypeptide. Polypeptide fused with His-tag was expressedat 37°C,for16hwith 1.0 mM IPTGinducer. Using Ni-NTA chromatographic column recombinant polypeptide of about 35 kDa from cell extractswas purified. The purified polypeptide will be used as antigenfor inducing specific antibody against outer capsid protein of SRBSDV.

  DOI: https://doi.org/10.31817/tckhnnvn.2015.13.7.