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鹅细小病毒NS2基因的原核表达及抗原性检测
Prokaryotic expression of goose parvovirus NS2 gene
【摘要】 为原核表达鹅细小病毒(GPV)NS2蛋白,本研究利用特异性引物扩增获得GPV的H1分离株NS2基因,将其克隆于pMD18-T载体后进行序列测定。并将NS2基因亚克隆于原核表达载体pGEX-6P-1中,获得重组质粒pGEX-NS2。该质粒转化于感受态菌BL21(DE3)plysS中,经IPTG诱导,SDS-PAGE电泳分析,表达的重组蛋白约为75ku左右。经过亲和层析方法获得了纯化的重组NS2蛋白。Westernblot和Dot-ELISA鉴定结果表明,表达的重组NS2蛋白可以与GPV阳性血清发生特异性反应。
【Abstract】 The goose parvovirus GPV NS2 gene was amplified by PCR and cloned into vector pGEX-6P-1. The recombinant plasmid pGEX-NS2 was transformed into BL21(DE3) pLysS E.coli and induced with IPTG. The expressed NS2 fusion protein had a MW of 75 ku and was purified by GST affinity chromatography purification system. Western blot and Dot-ELISA showed that the purified NS2 protein could be recognized by GPV positive serum.
【关键词】 鹅细小病毒;
NS2基因;
原核表达;
纯化;
抗原性;
【Key words】 GPV; NS2 gene; prokaryotic expression; purification; antigenicity;
【Key words】 GPV; NS2 gene; prokaryotic expression; purification; antigenicity;
【基金】 黑龙江“十五”攻关项目(GB01B503202)
- 【文献出处】 中国预防兽医学报 ,Chinese Journal of Preventive Veterinary Medicine , 编辑部邮箱 ,2010年02期
- 【分类号】S852.65
- 【被引频次】14
- 【下载频次】170