中国兽医科学2013，Vol.43Issue(7)：718-722,5.
犬新孢子虫AMA1基因重组真核表达质粒的构建及其在Vero细胞中的表达
Construction of recombinant eukaryotic expression plasmid of AMA1 gene of Neospora caninum and its expression in Vero cells
摘要
Abstract
To construct recombinant eukaryotic expression plasmid of AMA1 gene of Neospora canihum and express it in Vero cells,the full-length fragment of NcAMA1 gene from bovine N.caninum Jilin strain was amplified by RT-PCR,and then subcloned into the expression vector pVAX-1 to construct a eukaryotic expression plasmid pVAX1-AMA1.The recombinant was then transfected into the Vero cells by liposome method.The expressed product in Vero cells was analyzed by indirect immunofluorescence assay (IFA) and Western-blot.The results showed that the amplified fragment was 1 695 bp,and shared 99.9％ homology with the corresponding gene sequence in GenBank (AB265823.1).The recombinant eukaryotic expression plasmid pVAX1-AMA1 was successfully constructed.IFA result showed that the AMA1 gene was transiently expressed,and Western-blotting result showed that expressed product was 68 ku in molecular weight and had a positive reaction with anti-AMA1 serum.These results may provide a foundation for further studies on the development of nucleic acid vaccines against N.caninum infections.关键词
犬新孢子虫/AMA1基因/真核表达Key words
Neospora caninum / AMA1 gene/eukaryotic expression分类
农业科技引用本文复制引用
郭焕平,贾立军,薛书江,钱年超,张守发..犬新孢子虫AMA1基因重组真核表达质粒的构建及其在Vero细胞中的表达[J].中国兽医科学,2013,43(7):718-722,5.基金项目
国家自然科学基金资助项目(31160501) （31160501）
吉林省青年科研基金项目(201201076) （201201076）
吉林省自然科学基金项目(201115230) （201115230）
延边大学科技发展计划项目(延大科合字2011第37号) （延大科合字2011第37号）
博士科研启动基金(延大校发[2012]173号) （延大校发[2012]173号）
