微小牛蜱幼蜱cDNA表达文库的构建  

Construction of cDNA expression library from larvae of Boophilus microplus

作　　者：李文卉[1] 罗建勋[1] 殷宏[1] 高金亮[1] 关贵全[1] 刘志杰[1] 党志胜[1] 马米玲[1] 刘爱红[1] 任巧云[1] 

机构地区：[1]中国农业科学院兰州兽医研究所甘肃省动物寄生虫病重点实验室,甘肃兰州730046

出　　处：《中国兽医科技》2005年第12期994-996,共3页Chinese Journal of Veterinary Science and Technology

基　　金：国家高技术研究发展计划(863)项目(2003AA241110);国家自然科学基金与JSPS国际合作项目(30270992)

摘　　要：为了研究微小牛蜱功能性抗原基因,从微小牛蜱(Boophilus microplus)幼蜱中提取总RNA并分离纯化mRNA。采用Oligo(dT)引物合成双链cDNA,在其两端加EcoRⅠ、HindⅢ定向接头,用Mini Column Fractionation凝胶过滤柱纯化后定向克隆到λSCREEN载体。经体外包装,成功构建了我国微小牛蜱幼蜱cDNA表达文库,铺板测定原始库容量为4.5×105PFU,扩增后的滴度为7.2×1010PFU/mL。The total RNA and subsequently mRNA were isolated from the larvae of 13oophilus microplus to study functional genes of 13. microplus. （）ligo（dT） primed eDNA with directional EcoR Ⅰ/Hind Ⅲ linkers was synthesized and further purified by Mini Column Fractionation Kit, and then ligated to the EcoR Ⅰ/Hind Ⅲ arms of the λscreen vector. The eDNA expression library was constructed successfully with 4.5× 10^5PFU size of the primary library and 7.2 ×10^5PFU/mL titer of the amplified library.

关 键 词：微小牛蜱 CDNA表达文库 抗原基因 

分 类 号：S852.746[农业科学—基础兽医学] Q786[农业科学—兽医学]