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

Received: 26-03-2015

Accepted: 09-10-2015

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Issue: Vol. 13 No. 7 (2015)

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KỸ THUẬT VÀ CÔNG NGHỆ

How to Cite:

Hanh, D., Tam, P., & Hoi, P. (2024). Purification and Expression of DNA Segment Encoding Highly-Antigenic Polypeptide from Capsid Protein P10 of Southern Rice Black-Streaked Dwarf Virus. Vietnam Journal of Agricultural Sciences, 13(7), 1153–1161. http://testtapchi.vnua.edu.vn/index.php/vjasvn/article/view/1550

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

Do Thi Hanh (*) 1 , Pham Thanh Tam 2 , Pham Xuan Hoi 2

  • 1 Đại học Phương Đông
  • 2 Viện Di truyền Nông nghiệp

    • Keywords

    Highly-antigenic polipeptide, P10.1, recombinant protein, SRBSDV, pET28a expression vector

    Abstract


    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.

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