家畜生态学报2017，Vol.38Issue(10)：15-19,5.
长链非编码RNA AK005183干扰载体构建及慢病毒包装
Construction of the Plamid Carrying Short Hairpin RNA Targeting Long Noncoding RNA AK005183
摘要
Abstract
To study the function of long noncoding RNA AK005183 in sperms,the shRNA oligonucleotide interfering AK005183 expression was designed and synthesized,and its interference expression of recombinant lentiviral vector CD513B-U6-AK005183 was constructed as well.Transfected NIH3T3 cells with interference expression plasmid to detect the interference efficiency of this shRNA on the expression of AK005183 by real time and quantitive PCR.By using the third generation lentivirus package system,the plasmids of CD513B-U6-AK005183,pGag-Pol,pRsv-rev,and pVSV-G was transfected to 293T cell lines,an interference expression of recombinant lentivirus with titer of 5 × 106 TU/mL was aquired from the package.It was detected that the expression of AK005183 declined by 64％ due to the transfection of GC-lspa cells lines with lentivirus.This study successfully contructed and packaged the recombinant lentivirus which can highly and efficiently interfere with long noncoding RNA AK005183,providing a foudation for further research on function of long noncoding RNA in sperms.关键词
长链非编码RNA/AK005183/精原干细胞/RNA干扰/慢病毒Key words
long noncoding RNA/AK005183/Spermatogonial stem cells/RNA interfere/lentivirus分类
农业科技引用本文复制引用
张晓明,安俊辉,万一,胡源,覃金洲,曾文先..长链非编码RNA AK005183干扰载体构建及慢病毒包装[J].家畜生态学报,2017,38(10):15-19,5.基金项目
国家自然科学基金项目(31072029,31272439) （31072029,31272439）
