实时荧光定量PCR筛选牛源整联蛋白αv基因的siRNA片段  

Selection of siRNA Fragment of Cattle-borne Integrin-αv Gene by Real-time Fluorescence Quantitative PCR

作　　者：张国峰[1] 独军政[1] 常惠芸[1] 薛霜[1] 骆继怀[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部畜禽病毒学重点开放实验室国家口蹄疫参考实验室,甘肃兰州730046

出　　处：《动物医学进展》2011年第3期56-60,共5页Progress In Veterinary Medicine

基　　金：国家转基因重大专项(2008ZX08011-004;2009ZX08007-008B)

摘　　要：设计并合成三条针对牛口蹄疫病毒(FMDV)整联蛋白受体αv亚基基因的特异性干扰小RNA(SiRNA)片段,将其转染至本身表达整联蛋白受体αv亚基基因的MDBK细胞。分别在转染后36 h和48 h收集细胞,提取细胞总RNA并反转录为cDNA。应用SYBR GreenⅠ实时荧光定量RT-PCR的方法检测3个siRNA片段的干扰效果,筛选出对整联蛋白αv基因具有高效干扰效果的小RNA片段。为建立抗口蹄疫病毒牛新品种培育的RNA干扰转基因技术平台奠定了基础。In order to select the most efficient small interference RNA among three RNA oligoes which aim at downregulateing the expression of integrin-αv,the three RNA oligoes were transfected into the MDBK cells which express the integrin-αv gene.The cells were harvested in 36 hours and 48 hours respectively after transfection,and then the total RNA from these cells was extracted.After transcripting the RNA into cDNA by RT-PCR,these cDNA as templates carrying out fluorescence quantitative PCR were used to detect the efficience of the three siRNA.At last,the efficiency siRNA fragment of integrin-αv gene was selected.

关 键 词：实时荧光定量PCR 整联蛋白 RNA干扰 SYBR GreenⅠ 

分 类 号：Q786[生物学—分子生物学]