<FONT face="Verdana">Prokaryotic Expression of Coat Protein Gene  S10 of Rice BlackStreaked Dwarf Virus,  and Preparation and Application of Its Polyclonal Antibody </FONT>

OUYANG Yuan-long,WU Jian-xiang, XIONG Ru-yi,ZHOU Yi-jun,ZHOU Xue-ping

doi:10.3969/j.issn.1001-7216.2010.01.05

2010 , Vol. 24 >Issue 1: 25 - 30

DOI: https://doi.org/10.3969/j.issn.1001-7216.2010.01.05

研究报告

Prokaryotic Expression of Coat Protein Gene S10 of Rice BlackStreaked Dwarf Virus, and Preparation and Application of Its Polyclonal Antibody

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1College of Life Science, Nanjing Normal University, Nanjing 210046, China； 2Institute of Biotechnology， College of Agriculture & Biotechnology, Zhejiang University， Hangzhou 310029, China； 3Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China; *Corresponding author, E-mail: wujx@zju.edu.cn

Received date: 1900-01-01

Revised date: 1900-01-01

Online published: 2010-01-10

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Abstract

The full length cDNA of rice blackstreaked dwarf virus (RBSDV) segment 10 (S10) which encoded coat protein was cloned from the virus infected rice samples by RTPCR，and subcloned into a prokaryotic expression vector pET32a. The recombinant prokaryotic expression vector (pET32aCP) was used to transform Escherichia coli BL21 (DE3). A 76 kD TrxA fusion protein was obtained with induction of IPTG and purification of Ni+ NTA affinity column. The purified recombinant protein was used to immunize rabbits for production of polyclonal antibodies against the coat protein of RBSDV. Using polyclonal antibodies, immunocapture RTPCR and Dotblot ELISA were established for reliable, sensitive and specific detection of RBSDV. The two detection methods utilized polyclonal antibodies provide technical support for the diagnosis of RBSDV disease.

Key words： rice blackstreaked dwarf virus; prokaryotic expression; polyclonal antibody; immunocapture RTPCR; dotblot ELISA

Cite this article

OUYANG Yuan-long,WU Jian-xiang, XIONG Ru-yi,ZHOU Yi-jun,ZHOU Xue-ping . Prokaryotic Expression of Coat Protein Gene S10 of Rice BlackStreaked Dwarf Virus, and Preparation and Application of Its Polyclonal Antibody [J]. Chinese Journal OF Rice Science, 2010 , 24(1) : 25 -30 . DOI: 10.3969/j.issn.1001-7216.2010.01.05

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