CD146短发夹RNA真核表达载体的构建与鉴定  

Construction and identification of the eukaryotic expression vector expressing short hairpin RNA targeting human CD146

作　　者：陈伟[1] 董震[1] 蒋勇[1] 苏寒[1] 刘宝林[2] 孙沫逸[2] 

机构地区：[1]南京军区南京总医院口腔科,南京医学博士210002 [2]第四军医大学口腔医学院,西安710032

出　　处：《医学研究生学报》2011年第3期237-241,共5页Journal of Medical Postgraduates

基　　金：国家自然科学基金(30772428);南京军区南京总医院青年基金(2010Q009)

摘　　要：目的 CD146基因表达增高与涎腺腺样囊性癌(adenoid cystic carcinoma,ACC)嗜神经侵袭(perineural invasion,PNI)的临床症状相关。为进一步研究CD146基因的功能,构建人CD146短发夹RNA(short hairpin RNA,shRNA)真核表达载体并鉴定。方法根据Genebank中靶基因CD146序列,按照Tuschl设计原则,设计、合成编码其shRNA的互补寡核苷酸序列,克隆至线性化的pGenesil-1载体中并对重组质粒进行酶切分析及测序鉴定。结果经过限制性内切酶酶切分析和DNA测序鉴定,证实重组质粒pGe-CD146-shRNA的序列正确。结论成功构建了靶向人CD146基因的shRNA真核表达载体系统,为进一步研究CD146基因的功能奠定了实验基础。Objective The overexpression of CD146 is reportedly associated with perineural invasion in salivary adenoid cystic carcinoma.This study was designed to construct and identify the eukaryotic expression vector expressing short hairpin RNA（shRNA） targeting human CD146.MethodsTwo pairs of shRNA were designed according to the human CD146 sequence in the Genebank（NM_006500） following the rules of Tuschl,and then synthesized and cloned into the linearized pGenesil-1 vector.The recombinant vector was confirmed by enzyme digestion analysis and DNA sequencing.ResultsThe inserted sequence of the recombinant pGe-CD146-shRNA vector was exactly demonstrated by double restrictive endonuclease digestion and sequence analysis.ConclusionThe pGe-CD146-shRNA eukaryotic expression vectors were successfully constructed and identified.

关 键 词：CD146 短发夹RNA 真核表达载体 

分 类 号：Q782[生物学—分子生物学]