水貂肠炎病毒VP2基因原核表达及鉴定  

Prokaryotic Expression and Identification of Mink Enteritis Virus VP2 Gene

作　　者：林鹏[1] 王红梅[2] 王建科[1] 赵航[1] 郭利[1] 杨勇[1] 程悦宁[1] 张淼[1] 程世鹏[1] 

机构地区：[1]中国农业科学院特产研究所吉林省特种经济动物分子生物学重点实验室,长春130112 [2]山东省文登市畜牧兽医技术服务中心,文登264400

出　　处：《经济动物学报》2015年第3期133-139,共7页Journal of Economic Animal

基　　金：吉林省自然科学基金项目(20140101029JC);吉林省重点科技攻关项目(20150204021NY);吉林省特种经济动物生物制品科技创新中心;中国农业科学院科技创新工程(CAAS-ASTIP-2014-ISAPS)

摘　　要：采用PCR技术扩增MEV VP2基因,将该基因片段定向克隆原核表达载体PET-30a,构建MEV VP2基因原核表达重组质粒PET-30a-VP2。将该重组质粒转化到宿主菌BL21(DE3),IPTG诱导后进行SDSPAGE和Western-blotting分析。结果表明:该基因全长1 755 bp,其目的基因序列完全正确,能编码584个氨基酸。利用载体上的His标签对其进行纯化,经SDS-PAGE鉴定该重组蛋白以包涵体的形式存在,大小约为70 ku;Western-blotting验证该重组蛋白具有被抗MEV血清识别的特性,具有良好的反应原性,可为MEV基因工程亚单位疫苗、免疫学诊断方法等提供参考。The gene fragment of mink enteritis virus VP2 protein was amplified by PCR from MEV genome and cloned into PET-30a vector to construct recombinant prokaryotic expression plasmid PET-30a-VP2. The recombinant plasmid was transformed into Escherichia. coil BL21 （DE3） and ex- pression protein was analyzed by SDS-PAGE and Western-blotting. The result showed that the full length of VP2 gene included 1 755 nucleotides and encoded 584 amino acids, the target gene se- quence was completely correct. And the expression product was purified by His-tag of vector. SDS- PAGE analysis showed that the recombinant protein was about 70 ku and mainly expressed in the form of an inclusion body. Western-blotting analysis proved the recombinant protein had good reactive ability against MEV positive serum. The protein could be used for developing subunit vaccine and antigen of immunological diagnostic methods.

关 键 词：水貂肠炎病毒 VP2基因 原核表达 

分 类 号：S865.227[农业科学—野生动物驯养]