Author(s): ZHANG Yan-li, CAO Li-yan, LU Yun, GOU Hui-tian, WANG Yan-hua, FU Bao-quan, LI Wen-hui, ZHANG De-lin
Pages: 125-129
Year: 2009 Issue:  2
Journal: Veterinary Science in China
Keyword:  刚地弓形虫; 微线体蛋白3; 基因克隆; 真核表达;
Abstract: 采用PCR技术从刚地弓形虫GJS株基因组DNA中扩增微线体蛋白3(MIC3)基因,并克隆到pMD18-T载体,经PCR、酶切及测序鉴定后,阳性重组质粒酶切并亚克隆到真核表达载体pcDNA3.1(+)后进行PCR、酶切及测序鉴定.将重组质粒转染BHK-21细胞.间接免疫荧光染色证明,质粒DNA成功转染到细胞中并得以表达.Western-blot分析证实,细胞裂解液及上清样品中有1条约39.2 ku的条带,可被山羊抗刚地弓形虫超免疫血清所识别,大小与预测值相符.表明,真核表达质粒pcDNA3-MIC3中的MIC3基因在BHK-21细胞中获得表达且表达产物具有抗原性.