新孢子虫与弓形虫交叉抗原基因AMA1的克隆及原核表达载体构建  

Clone and prokaryotic expression vector construction of Neospora caninum and Toxoplasma gondii AMA1 gene

作　　者：孙婷婷[1] 金京顺 贾立军[1] 

机构地区：[1]延边大学农学院 [2]延吉市朝阳川镇畜牧兽医站,吉林延吉133000

出　　处：《延边大学农学学报》2015年第1期8-11,共4页Agricultural Science Journal of Yanbian University

基　　金：国家自然科学基金(31160501;31360605);吉林省重点科技攻关项目(20140204078NY);吉林省青年科研基金(201201076);国家级和吉林省"大学生创新创业训练计划项目"(教高司函[2014]58号1878;吉教高字[2014]27号453)

摘　　要：为了解新孢子虫与弓形虫交叉抗原基因AMA1的生物学特性,应用PCR技术扩增新孢子虫/弓形虫AMA1基因,将PCR产物纯化回收后克隆于pMD18-T Simple Vector,构建pMD18-T-Nc/Tg AMA1重组克隆质粒,经PCR鉴定、酶切鉴定及测序分析后,亚克隆于原核表达载体pGEX-4T-1,构建重组原核表达质粒pGEX-4T-Nc/Tg AMA1,经PCR鉴定、酶切鉴定表明,构建的原核表达载体正确。该试验为新孢子虫/弓形虫AMA1基因重组疫苗的研究奠定了基础。In order to understand the biological characteristics of Neospora and Toxoplasma＇ s antigen gene AMAI,we was amplified Neospora/Toxoplasma AMA1 gene by PCR, purified the product of PCR and connected to the cloning vector pMD18-T simple vector to con- struct pMD18 -T-Nc/Tg AMA1 recombinant plasmid clones. The expression plasmid which is identifieated correctly by PCR and enzyme digestion and sequencing analysis will be connected to prokaryotic expression vector pGEX-4T-1, eventually build prokaryotic expression vector pGEX-4T-Nc/Tg AMA1. It will provide a foundation for the study of Neospora/Toxoplasma AMA 1 recombinant vaccine.

关 键 词：新孢子虫 弓形虫 AMA1 克隆 PGEX-4T-Nc/Tg AMA1 

分 类 号：S855.9[农业科学—临床兽医学]