鹅细小病毒VP2基因在大肠杆菌中的表达及纯化  

Expression of Goose Parvovirus VP2 Protein in E.coli and Its Purification

作　　者：贺云霞[1] 王君伟[2] 王立群[1] Ulrich Neumann 

机构地区：[1]东北农业大学生命科学院,黑龙江哈尔滨150030 [2]东北农业大学动物医学院,黑龙江哈尔滨150030 [3]汉诺威兽医学院,德国汉诺威30559

出　　处：《中国兽医学报》2004年第2期126-128,共3页Chinese Journal of Veterinary Science

基　　金：黑龙江省"十五"攻关项目 (GB0 1B5 0 3 -0 2 )

摘　　要：将鹅细小病毒 (GPV) VP2基因插入原核表达载体 p PROEX- HTb,获得重组表达质粒 p PROEX- HTb- VP2。将其转化大肠杆菌 DH5α,用 IPTG诱导表达 ,表达菌体蛋白中可产生与预期大小相符的约 72 0 0 0的蛋白。以光密度扫描对该表达产物进行定量分析 ,表达产物约占菌体总蛋白的 14 %。经 His- tag金属螯合层析纯化 ,获得纯度较高的 6 His- VP2融合蛋白。 Western- blot结果表明 ,所得蛋白与鹅抗 GPV高免血清有较好的免疫反应性 ,说明在 VP2的 N端融合 6个组氨酸不影响其与特异抗体结合的活性。By inserting the goose parvovirus VP2 gene into prokaryotic expression vector pP_(ROEX)-HTb,the recombinant plasmid pP_(ROEX)-HTb-VP2 was constructed.Then it was transformed into E.coli DH5α and induced with IPTG.SDS-PAGE analysis showed a protein about (72 000) its molecular weight being consistent with GPV-VP2 protein was expressed.Densitometric scanning indicated that the expressed protein was 14% of total protein of DH5α.The protein was purified by metal(Ni^(2+)) chelation affinity chromatography and tested by Western-blot.It revealed that 6His-VP2 fusion protein has a positive reaction with anti-GPV serum,suggesting that the 6His-tag did not affect the specific antibody-recognizing activity of VP2 protein.

关 键 词：鹅细小病毒 VP2基因 原核表达 纯化 

分 类 号：S852.65[农业科学—基础兽医学] Q78[农业科学—兽医学]