牛bta-miR-26b靶基因EphA2的鉴定  

Identification of EphA2 as a novel target of bta-miR-26b in bovine

作　　者：于汪洋[1,2] 李凤霞[3] 孙喆[4] 袁宝[2] 高妍[2] 马腾壑[1,2] 戴立胜[1,2] 徐靖博[1,2] 孙广杰[1,2] 许橙[1] 张嘉保[2] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]吉林大学实验动物中心,吉林长春130062 [3]吉林大学农学部图书馆,吉林长春130062 [4]吉林农业大学动物科技学院,吉林长春130118

出　　处：《中国兽医学报》2012年第4期566-569,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30972100);“863”子课题资助项目(2008AA101010)

摘　　要：利用miRNA靶基因预测数据库TargetScan,RNA22对bta-miR-26b的靶基因进行预测,并鉴定bta-miR-26b的靶基因。生物信息学分析结果显示,在EphA2基因3′非编码区域(3′-UTR)有bta-miR-26b的保守结合种子序列。扩增含有bta-miR-26b结合位点的靶基因的3′-UTR区,插入双荧光素酶报告载体pmirGLo Dual-LuciferasemiRNA Target Expression vector,与bta-miR-26bmimics共转染Hela细胞后检测荧光素酶的活性变化。通过荧光素酶活性检测证明在体外bta-miR-26b可以与EphA2基因结合,证明EphA2可能为bta-miR-26b的一个靶基因。The potential target of bta-miR-26b was predicted by using 2 currently available prediction programs,including TargetScan and RNA22.EphA2 was chosen because it was predicted by all the 2 programs.The 3′-UTR sequence of EphA2 was obtained by chemical synthesis.The prediction results show that EphA2 is a potential target gene of miR-26b.The targeting of miR-26b to the 3′-UTR of the EphA2 was done using the pmirGLo Dual-Luciferase miRNA Target Expression vector,co-transfection of the vector with miR-26b mimics in Hela cells was analyed by detecting the luciferase activity.Luciferase reporter experiment shows that miR-26b could specifically bind to the EphA2 3′-UTR to inhibit the luciferase expression level.It was proved that miR-26b could target to EphA2 at mRNA level.

关 键 词：miR-26b EPHA2 荧光素酶活性 

分 类 号：S852.23[农业科学—基础兽医学]