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茎部长度为21、27、29bp的shRNA表达载体构建
引用本文:李莉,刘西梅,郑新民,刘中华.茎部长度为21、27、29bp的shRNA表达载体构建[J].湖北农业科学,2010,49(9).
作者姓名:李莉  刘西梅  郑新民  刘中华
作者单位:1. 湖北省农业科学院畜牧兽医研究所/动物胚胎工程及分子育种湖北省重点实验室,武汉,430064
2. 西北农林科技大学动物医学学院,陕西,杨凌,712100
基金项目:国家自然科学基金项目,动物胚胎工程及分子育种湖北省重点实验室项目 
摘    要:利用shRNA(Small hairpin RNA)载体调控基因表达已经成为研究基因功能的有力工具。针对增强型绿色荧光蛋白标记基因(Enhanced green fluorescent protein,egfp),分别设计茎部长度为21bp、27bp、29bp的干扰片段,体外合成的单链shRNA片段退火后连入带有U6启动子的真核表达载体psiSTRIKE中,酶切及测序结果证明设计的干扰片段已经成功克隆进入表达载体,为进一步在小鼠细胞以及个体水平上综合评价不同茎部长度shRNA的干扰效应奠定了基础。

关 键 词:茎部长度  小发夹RNA  基因沉默  RNA干扰  绿色荧光蛋白基因

Construction of shRNA Expression Vectors Embracing 21 bp,27 bp,or 29 bp Stem
LI Li,LIU Xi-mei,ZHENG Xin-min,LIU Zhong-hua.Construction of shRNA Expression Vectors Embracing 21 bp,27 bp,or 29 bp Stem[J].Hubei Agricultural Sciences,2010,49(9).
Authors:LI Li  LIU Xi-mei  ZHENG Xin-min  LIU Zhong-hua
Institution:LI Li1,LIU Xi-mei1,ZHENG Xin-min1,LIU Zhong-hua2(1.Institute of Veterinary and Animal Science,Hubei Academy of Agriculture Science/Hubei Key Lab of Animal Embryo & Molecular Breeding,Wuhan 430064,China,2.College of Veterinary Medicine,Northwest A & F University,Yangling 712100,Shaanxi,China)
Abstract:The use of small hairpin RNA(shRNA) to regulate gene expression is evolving into an increasingly powerful tool for gene function study in organisms.We designed shRNAs with stems of 21 bp,27 bp,and 29 bp targeting to the enhanced green fluorescent protein gene.The synthesized single strands were annealed and cloned into psiSTRIKE including U6 promoter.Interfering fragments had successfully been cloned into expression vector tested by enzyme digesting and sequencing.The work in this study will help us to valu...
Keywords:stem length  shRNA  gene silence  RNA interference  green fluorescent protein gene  
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