南芥菜花叶病毒外壳蛋白的克隆表达及抗血清的制备  

Cloning and Expression the cp Gene of ArMV in E.coli BL21 and Preparation Its Antiserum

作　　者：薛杨[1] 安德荣[1] 全鑫[1] 李明福[2] 

机构地区：[1]西北农林科技大学植保学院与陕西省农业分子生物学重点实验室,陕西杨凌712100 [2]中国检验检疫科学研究院动植物检疫研究所,北京朝阳100029

出　　处：《西北农业学报》2006年第6期217-220,共4页Acta Agriculturae Boreali-occidentalis Sinica

摘　　要：依据报道的ArMV外壳蛋白基因(coat protein,cp)序列设计引物,通过RT-PCR扩增得到长约990bp的ArMV cp基因亲水基团部分片段,将目的片断克隆到原核表达载体pEGX-6p-1中,构建ArMV cp基因与GST蛋白融合表达载体pEGX-cp,重组载体化大肠杆菌BL21,经IPITG诱导后,融合蛋白GST-cp得到了特异表达。用12%的SDS-PAGE分析结果表明,表达产物大小为63.2 kDa主要以包含体的形式存在。用冰冷的KCl显色,分离表达的蛋白质特异条带制备抗体,免疫家兔后得到特异性较强的抗血清,经酶联免疫法检测效价为1∶100 000。A pair of primers was designed according to the coat protein （cp） gene of Arabis mosaic viru （ArMV） and used to amplify a 990 bp partial gene of the cp by reverse transcription-polymerase chain reaction （RT-PCR）, the 990 bp partial gene was cloned into the pEGX-9p-1, a prokaryotic expression vector fused with GST. The recombinant vector was transformed into E. coli BL21. The fusion protein was successfully expressed in E. coli BL21 induced with the IPTG at 37℃. 12% SDS-PAGE showed that the expressed product with molecular weight 63.2kDa was mainly in the form of inclusion body. Rabbit was immunized with the expressed fusion protein as antigen and abtiserum was obtained. And the titer was determined to be 1 ： 100 000 by ELISA.

关 键 词：南芥菜花叶病毒 外壳蛋白 融合蛋白 抗血清 

分 类 号：S432.41[农业科学—植物病理学]