靶向口蹄疫病毒shRNA表达载体转染后特异性siRNA的分子检测  

Detection of specific siRNA after transfection of shRNA expression vector targeting FMDV

作　　者：蔡扩军[1] 孟庆玲[1] 乔军[1] 陈创夫[1] 马饰委[1] 黄炯[2] 张再超[1] 田振中[1] 杨丽红[1] 

机构地区：[1]石河子大学动物科技学院,新疆石河子832003 [2]新疆畜牧科学院兽医研究所,新疆乌鲁木齐830000

出　　处：《中国兽医学报》2013年第1期49-52,共4页Chinese Journal of Veterinary Science

基　　金：国家转基因重大专项(2009ZX08006-003B)

摘　　要：通过Poly A polymerase对小干扰RNA分子(Small interfering RNA,siRNA)3′端进行加"Poly A"处理,然后进行RT-PCR扩增,并将扩增产物克隆到T载体上进行测序。结果表明,通过该方法可以快速检测FMDV特异性siRNA分子的表达,这为siRNA的干扰FMDV复制分子机制和抗FMDV转基因动物细胞水平的评价研究提供了技术支撑。RNA interference,mediated by small interfering RNA,is a fast developed post-transcriptional gene silencing method in resent years,which has been extensively used in the fields of gene function research.To identify wether siRNA was expressed in BHK-21 cells after interference vector targeting Foot-and-mouth disease（FMDV） was transfected,Poly A polymerase was used to add poly-A tails to the 3′ end of siRNA,then reverse transcription PCR（RT-PCR） technique was and introduced to detect siRNA expression,the product was subcloned into T Vector and sequenced.Experiment results showed that this method is a fast and facile tool to detect siRNA and study siRNA interfering mechanisms in FMDV replication and assessment of transgenic animal cells anti-FMDV.

关 键 词：SIRNA 表达 反转录PCR 口蹄疫病毒 

分 类 号：S852.652[农业科学—基础兽医学]