摘要
以猪瘟病毒全长基因组质粒为模板,PCR扩增出E2囊膜糖蛋白上A/D抗原区B细胞表位878aa～938aa;PCR扩增片段连接到pET-32a(+)载体构建pET-AD原核表达质粒,将阳性质粒转化到BL21大肠埃希菌中诱导表达,以Ni-NTA亲和柱纯化,纯化后的蛋白进行Western blot鉴定。结果表明,重组质粒pET-AD在BL21大肠埃希菌中可以高效表达,表达的蛋白质经纯化后仍然有反应原性。
Using CSFV genome plasmid as PCR template, the specific gene encoding the aa878-aa938 A/D antigenic domains in E2 protein was amplified. The PCR product was digested with EcoR Ⅰ and Hind Ⅲ and inserted into pET-32a （＋） to get pET-AD recombinant plasmid. The positive recombinant vectors were transformed into the competent cells, BL21 to get recombinant bacteria. The recombinant bacteria were induced with IPTG and the obtained protein were purified by Ni-NTA affinity chromatography. Purified protein was analyzed by Western blot. The results showed that the recombinant plasmid pET-AD could express in BL21 with high level, and the expressed product could react with antibody effectively.
出处
《动物医学进展》
CSCD
2008年第5期40-42,共3页
Progress In Veterinary Medicine
关键词
猪瘟病毒
抗原表位
表达
免疫印迹
Classical swine fever virus
epitope
expression
Western blot
