弓形虫RH株表面抗原SAG3去信号肽基因的蛋白原核表达及鉴定  

Protein Prokaryotic Expression and Identification of Excluding Signal Peptide Gene in SAG3 of Toxoplasma RH Strain

作　　者：马亮[1,2] 刁玉梅[1] 任保彦[1] 宫鹏涛[1] 李建华[1] 张西臣[1] 

机构地区：[1]吉林大学畜牧兽医学院,长春130062 [2]吉林市动物检疫站,吉林132013

出　　处：《吉林农业大学学报》2012年第1期104-108,共5页Journal of Jilin Agricultural University

基　　金：国家科技支撑计划项目(2008BAD96B11-3)

摘　　要：根据GenBank中弓形虫表面抗原SAG3基因序列,以弓形虫总RNA反转录的cDNA为模板,扩增出SAG3去除信号肽基因并进行原核表达。将重组pET-28a(+)-SAG3阳性表达质粒转入大肠杆菌BL(DE3)中,用IPTG进行诱导表达。通过SDS-PAGE和Western blotting对重组蛋白进行分析和鉴定。结果表明:成功扩增了不含信号肽的SAG3基因,构建的原核表达质粒在大肠杆菌中得到了高效表达,能够与鼠抗弓形虫阳性血清发生特异性反应,去信号肽的SAG蛋白具有反应原性。The research was conducted to clone, express and identify the SAG3 gene containing no signal peptide of Toxoplasma gondii. The SAG3 gene containing no signal peptide was amplified by RTPCR. The prokaryotic expression plasmid pET-28a（ ＋ ）-SAG3 was constructed by sub-cloning the SAG fragment into the prokaryotic expression vector pET-28a（ ＋ ）. The expression of pET-28a-SAG3 was induced by IPTG in E. coli BI21 （DE3）system; then the fusion protein was identified by SDS - PAGE and Western blotting. The results indicated that the fusion protein was expressed in E. coli BI21 （DE3） and could be specially recognized by polyclonal antibody against the SAG3 of Toxoplasma gondii. The results were useful for further studies on the diagnosis of toxoplasmosis by SAG3 gene containing no signal peptide.

关 键 词：刚地弓形虫 SAG3基因 原核表达 

分 类 号：S852.72[农业科学—基础兽医学]