中国动物传染病学报2012，Vol.20Issue(5)：39-44,6.
柔嫩艾美耳球虫棒状体颈部蛋白2基因真核表达载体的构建及在293T细胞中表达
CONSTRUCTION OF EUKARYOTIC EXPRESSION VECTOR OF EIMERIA TENELLA RHOPTRY NECK PROTEIN 2 GENE AND ITS EXPRESSION IN 293T CELLS
摘要
Abstract
To construct an eukaryotic expression vector for Eimeria tenella rhoptry neck protein 2 (EtRON2) gene and study the expresstion of EtRON2 gene in transfected 293T cells, a part of EtRON2 gene was amplified by PCR from the cDNA of Eimeria tenella sporulated oocyst and cloned into PGEM-Teasy vector to construct a vector named pGEM-Teasy-EtRON2. The pGEM-Teasy-EtRON2 and the pCAGGS vector were digested by the same restriction enzymes and the recombinant eukaryotic expression vector pCAGGS- EtRON2 was subsequently constructed. The recombinant plasmid pCAI3GS-EtRON2 was identified and transfectcd into 293T cells. The expression of EtRON2 protein was verified by indirect immunofluorescence assay and Western blot. Restriction enzyme digestion obtained correct length ofEtRON2 gene of about 1172 bp. Sequence analysis confirmed EtRON2 gene sequence that was exactly the same as that deposited in GenBank. The results of Western blot showed that the molecular weight of expressed protein was around 43 kDain accordance with the target protein. Red fluorescence was observed in indirect immunofluorescence assay. Taken together, the eukaryotic expression vector of pCAGGS-EtRON2 was constructed and expressed in 293T cells, which would establish the foundation for the future research on its biological functions.关键词
柔嫩艾美耳球虫/棒状体颈部蛋白2/293T细胞/真核表达Key words
Eimeria tenella/rhoptry neck protein 2/293T cell/erkaryotic expression分类
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王晔,姜连连,韩红玉,薛璞,赵其平,董辉,朱顺海,舒凡帆,黄兵..柔嫩艾美耳球虫棒状体颈部蛋白2基因真核表达载体的构建及在293T细胞中表达[J].中国动物传染病学报,2012,20(5):39-44,6.基金项目
中央级公益性科研院所基本科研业务费项目 （）
