猪繁殖与呼吸综合征病毒核衣壳蛋白基因的原核表达及纯化  

Prokaryotic Expression and Purification of Nucleocapsid Protein of Porcine Reproductive and Respiratory Syndrome Virus

作　　者：于丽[1] 邵烈刚[1] 刘海军[1] 胡仁建[1] 余华[1] 叶芬[1] 蔡家利[1] 

机构地区：[1]西南大学动物科技学院,重庆400715

出　　处：《动物医学进展》2010年第12期58-61,共4页Progress In Veterinary Medicine

摘　　要：利用含有PRRSV N蛋白基因的重组表达质粒pREST-N的大肠埃希菌,在37℃条件下,用终浓度为1 mmol/L的IPTG进行诱导表达,经SDS-PAGE分析,目的基因高效表达出分子质量约为17.8 ku的可溶性融合蛋白,分子质量与预期设计的N蛋白大小相符。经His-Ni2+柱亲和层析纯化后,得到了高纯度的N蛋白。通过Western blot分析,该重组N蛋白与兔抗PRRSV高免血清能够发生特异性反应,说明其具有良好的反应原性,为进一步研究PRRSV的抗体检测方法及诊断抗原的批量生产工艺奠定了基础。N protein gene of PRRSV was cloned into the pRSET A vector to construct a recombinant expression plasmid pREST-N,which was transformed into E.coli BL21.The engineering bacteria were induced by IPTG of 1 mmol/L at 37 ℃.The molecular weight of recombinant N protein,in the form of soluble fusion body,was approximately 17.8 ku,consistent with that of the designed.The expressed N protein was purified by His-Ni2＋ affinity chromatography and high purity was obtained.Analyzed by Western-blot,the N protein can react with the rabbit anti-PRRSV positive serum,which indicated it possessed good antigenicity.This assay layed foundation for the further research of PRRSV antibody test and the mass production of diagnostic antigen.

关 键 词：猪繁殖与呼吸综合征病毒 N蛋白 表达质粒pREST-N 纯化 

分 类 号：Q786[生物学—分子生物学] S852.659.6[农业科学—基础兽医学]