增强型绿色荧光蛋白RNAi表达载体的构建和鉴定  

Construction and identification of RNA interference expression vector targeting EGFP

作　　者：干惠珠[1] 张桂珍[2] 卜丽莎[2] 杨绍娟[2] 高申 郑德明[2] 

机构地区：[1]吉林大学中日联谊医院肿瘤血液科,吉林长春130033 [2]吉林大学中日联谊医院中心研究室

出　　处：《中国实验诊断学》2011年第4期619-621,共3页Chinese Journal of Laboratory Diagnosis

摘　　要：目的构建靶向增强型绿色荧光蛋白基因(EGFP)的RNA干扰质粒载体。方法设计并合成针对EGFP编码区的模板寡核苷酸单链,克隆入pSilencerTM 3.1-H1 neo载体中,对重组质粒进行酶切鉴定和DNA序列测定。结果重组质粒经限制性内切酶酶切得到66 bp和4.3 kb条带;重组阳性克隆测序结果与实验设计的模板寡核苷酸序列相符。结论成功构建靶向EGFP的RNA干扰表达载体,为RNA干扰技术在实验室的进一步开展奠定基础。Objective To construct RNAi（RNA interference） expression vector targeting EGFP.Methods The oligos targeting EGFP were desigened and cloned into the BamHⅠand Hind Ⅲ site of linearized pSilencerTM 3.1-H1 neo shRNA expression vector.The recombinant plasmid vectors were confirmed by enzyme digestion analysis and DNA sequencing.Results The fragment of 66bp and 4.3kb was shown after digestion by BamHⅠ和Hind Ⅲ and agarose gel electrophoresis.,which was proved to be identical to original construction.Conclusion RNAi expression vector targeting EGFP has been constructed successfully,which facilitates the wider use of RNA interference technique in laboratory.

关 键 词：RNAI 增强型绿色荧光蛋白 SHRNA表达载体 

分 类 号：R346[医药卫生—基础医学]