应用RNAi干扰eGFP转基因小鼠胎儿成纤维细胞中FGF5基因的表达  

Suppression of FGF5 in Fetal Fibroblasts Derived from eGFP Transgenic Mice by RNA Interference

作　　者：宝明涛[1] 郭旭东[1] 旭日干[1] 

机构地区：[1]内蒙古大学实验动物研究中心教育部重点实验室,呼和浩特010021

出　　处：《四川动物》2008年第5期785-787,共3页Sichuan Journal of Zoology

基　　金：国家863计划项目(2002AA242061);内蒙古自然科学基金(200607010405);内蒙古自治区高等学校科学研究项目(NJ06055)资助

摘　　要：在小鼠FGF5基因干扰的研究中,针对小鼠FGF5mRNA的第316～335bp区域、第499～518bp区域和第766～785bp区域分别设计了发夹式RNA干扰片段,并将干扰片段连接到带有H1启动子的红色荧光表达载体上,将载体以脂质体法转染到eGFP转基因小鼠胎儿成纤维细胞中,搜集转染后的细胞提取总RNA,并反转录成cDNA。用SYBRGREENⅠ荧光定量PCR方法对转染了不同干扰载体的细胞cDNA进行检测,结果干扰载体对eGFP转基因小鼠成纤维细胞中的FGF5表达有较强的抑制作用。Three short hairpin RNA （shRNA） vectors targeting three different regions （ 316 - 335 bp,499 - 518 bp and 766 - 785 bp） of fibroblast growth factor 5 （ FGF5 ） mRNA were designed and constructed under the control of a red fluorenscent protein expression cassette with H1 promoter. After that the shRNA vectors were transfeeted into the eGFP cells derived from fetal eGFP transgenic mice, respectively, and the total RNA were extracted, then reversed transcripted to eDNA. Subsequently the eDNA are detected by the method of SYBR GREEN Ⅰ Real Time PCR. The conclusion is that the shRNA vectors can knockdown the expression of FGF5 gene in fetal fibroblasts derived from eGFP transgenie mice.

关 键 词：RNAI SHRNA SYBR GREEN Ⅰ荧光定量PCR FGF5 

分 类 号：Q78[生物学—分子生物学]