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当前位置: 首页 > 文章 > 绵羊细粒棘球蚴Ag B基因的克隆、表达及其重组蛋白纯化 石河子大学学报(自然科学版) 2014,32 (6) 687-691
Position: Home > Articles > Cloning,Expression of Ag B Gene from Echinococcus granulosus and Purification of Recombinant Protein Journal of Shihezi University(Natural Science) 2014,32 (6) 687-691

绵羊细粒棘球蚴Ag B基因的克隆、表达及其重组蛋白纯化

作  者：
刘田莉;易继海;营瑞文;宋雪梅;乔军;孟庆玲;胡政香;才学鹏;陈创夫
单  位：
石河子大学动物科技学院;中国农业科学院兰州兽医研究所
关键词：
细粒棘球蚴;Ag B基因;原核表达;绵羊
摘  要：
为克隆、表达绵羊细粒棘球蚴Eg Ag B基因,并纯化重组蛋白,采用PCR方法从绵羊肝脏包囊病料中扩增Ag B基因,克隆入p MD18-T载体中,测序后进行序列分析。将Ag B基因亚克隆入原核表达质粒p ET-28a中,转化入E.coli BL21感受态细胞中,用IPTG诱导Ag B蛋白表达,并对重组蛋白进行SDS-PAGE和Western blot分析。结果显示:克隆的Ag B基因全长333 bp,通过SDS-PAGE可以检测到分子量为20 ku的特异性重组蛋白,与预期值相符;Western blot分析结果显示,该蛋白可与Eg阳性血清发生免疫学反应。本研究在大肠杆菌中成功表达的绵羊细粒棘球蚴Ag B蛋白,并证实其具有良好的反应原性,为绵羊细粒棘球蚴感染检测试剂的研发奠定了基础。
译  名：
Cloning,Expression of Ag B Gene from Echinococcus granulosus and Purification of Recombinant Protein
作  者：
LIU Tianli;YI Jihai;YING Ruiwen;SONG Xuemei;QIAO Jun;MENG Qingling;HU Zhengxiang;CAI Xuepeng;CHEN Chuangfu;College of Animal Science and Technology,Shihezi University;Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences;
关键词：
Echinococcus granulosus;;Ag B;;prokaryotic expression;;sheep
摘  要：
In order to clone,express the Ag B gene from sheep Echinococcus granulosus(Eg) and purify the recombinant protein,the Ag B gene was amplified from the infected cyst disease of sheep liver by PCR method.Then it was cloned into p MD 18-T vector for sequence analysis.The Ag B gene was subcloned into the prokaryotic expression vector of p ET-28 a,and transformed into E.coli BL21 competent cells,and induced with IPTG for Ag B protein expression.The recombinant protein was analyzed by SDS-PAGE and Western blot.The results showed the nucleotide of Ag B gene had a length of 333 bp.The specific 20 ku fusion protein was examined by SDS-PAGE,which is consistent with expectations.Western blot analysis showed that the protein has the immune response with the positive serum against Eg.Ag B protein of Eg was successfully expressed in E.coli in the study,and confirmed its good reactogenicity,which laid the foundation for the development of detection reagent of sheep infected by Eg.

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