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  

Polyclonal antibody preparation against prokaryotic expression products of Rice stripe virus NS2 gene and detection of NS2 in infected rice and planthoppers

作　　者：熊如意[1,2] 吴建祥[1] 周益军[2] 周雪平[1] 

机构地区：[1]浙江大学生物技术研究所,杭州310029 [2]江苏省农业科学院植物保护所,南京210014

出　　处：《植物病理学报》2009年第1期95-99,共5页Acta Phytopathologica Sinica

基　　金：浙江省自然科学基金资助项目(Y305013);江苏省自然科学基金资助项目(BK2006164);农业部行业专项(NYHYZX07-01)

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.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.

关 键 词：水稻条纹病毒 NS2基因 灰飞虱 抗体制备 表达产物 多克隆 水稻条纹叶枯病 检测 

分 类 号：S435.111.4[农业科学—农业昆虫与害虫防治] S852.659.2[农业科学—植物保护]