Dnd1基因RNAi慢病毒载体的构建与鉴定  

Construction and Identification of the shRNA lentiviral expression Vector for targeting mouse Dnd1 gene

作　　者：李顺东 于淼[2] 张勇[2] 邓来[3] 谭宇婷[2] 程洁 彭小宁[2] 

机构地区：[1]长沙市第三医院,湖南长沙410005 [2]湖南师范大学医学院 [3]中南大学湘雅二医院,湖南长沙410006

出　　处：《湖南师范大学学报（医学版）》2011年第3期12-15,共4页Journal of Hunan Normal University(Medical Sciences)

基　　金：长沙市科技计划资助项目(k0802084-31)

摘　　要：目的:构建小鼠Dnd1基因的RNAi慢病毒载体。方法:针对小鼠Dnd1基因RNAi设计有效靶序列,合成靶序列的Oligo DNA。退火形成双链DNA,与经Age I和EcoR I酶切后的pGCSIL-GFP载体连接产生短发卡RNA慢病毒载体,PCR筛选阳性克隆,测序鉴定。结果:PCR鉴定与DNA测序证实合成的含Dnd1sh RNA慢病毒载体寡核苷酸链插入正确。结论:成功构建小鼠Dnd1基因的RNAi慢病毒载体。Objective To construct the shRNA lentiviral expression vector for targeting mouse Dnd1 gene.Methods Four pairs of sh RNAs that target at Dnd1 gene were designed.After annealed,the inserts were ligated into the linearized pGCSIL-GFP,and the vectors were identified by PCR and DNA sequencing.Results The target segments were cloned into pGCSIL-GFP lentivirus RNAi vector respectively,and the recombinant plasmids containing Dnd1 sh RNA was confirmed correct by PCR identification and DNA sequencing.Conclusion We successfully constructed four recombinant plasmids pGCSIL-GFP-sh RNA that target at Dnd1 gene,and it can establish a foundation for our study the molecular function of Dnd1.

关 键 词：Dnd1基因 慢病毒载体 RNA干扰 

分 类 号：R346[医药卫生—基础医学]