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当前位置: 首页 > 文章 > 水稻条纹病毒NS2基因原核表达产物多克隆抗体制备及受侵染水稻和灰飞虱体内NS2检测 植物病理学报 2009,39 (1) 95-99
Position: Home > Articles > Polyclonal antibody preparation against prokaryotic expression products of Rice stripe virus NS2 gene and detection of NS2 in infected rice and planthoppers Acta Phytopathologica Sinica 2009,39 (1) 95-99

水稻条纹病毒NS2基因原核表达产物多克隆抗体制备及受侵染水稻和灰飞虱体内NS2检测

作  者：
熊如意;吴建祥;周益军;周雪平
单  位：
江苏省农业科学院植物保护所;浙江大学生物技术研究所
摘  要：
The NS2 gene of Rice stripe virus(RSV) was amplified by RT-PCR,cloned into pGEM-T vector and sequenced.The NS2 gene was inserted into prokaryotic expression vector pET32a to produce recombinant plasmid pET32a-NS2.The recombinant plasmid was introduced into Escherichia coli strain BL21(DE3) pLysS.SDS-PAGE and Western blot analysis confirmed that NS2 fusion protein was expressed after induction by IPTG.The recombinant NS2 protein was purified with Ni2+-NTA agarose affinity chromatography and the polyclonal antibody against NS2 protein was raised in rabbit.NS2 protein was successfully detected in small brown planthopper(Laodelphax striatellus) at 1∶1 600 dilution of the total protein of single planthopper and in infected rice(Oryza sativa) at 1∶800 dilution of 10 mg leave by dot immunobinding assay using the polyclonal antibody.
译  名：
Polyclonal antibody preparation against prokaryotic expression products of Rice stripe virus NS2 gene and detection of NS2 in infected rice and planthoppers
作  者：
XIONG Ru-yi1,2,WU Jian-xiang1,ZHOU Yi-jun2,ZHOU Xue-ping1 (1Institute of Biotechnology,Zhejiang University,Hangzhou 310029,China;2Institute of Plant Protection,Jiangsu Academy of Agricultural Science,Nanjing 210014,China)
关键词：
Rice stripe virus;NS2 gene;prokaryotic expression;preparation of polyclonal antibody;detection
摘  要：
The NS2 gene of Rice stripe virus(RSV) was amplified by RT-PCR,cloned into pGEM-T vector and sequenced.The NS2 gene was inserted into prokaryotic expression vector pET32a to produce recombinant plasmid pET32a-NS2.The recombinant plasmid was introduced into Escherichia coli strain BL21(DE3) pLysS.SDS-PAGE and Western blot analysis confirmed that NS2 fusion protein was expressed after induction by IPTG.The recombinant NS2 protein was purified with Ni2+-NTA agarose affinity chromatography and the polyclonal antibody against NS2 protein was raised in rabbit.NS2 protein was successfully detected in small brown planthopper(Laodelphax striatellus) at 1∶1 600 dilution of the total protein of single planthopper and in infected rice(Oryza sativa) at 1∶800 dilution of 10 mg leave by dot immunobinding assay using the polyclonal antibody.

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