雌激素受体ERbeta的克隆及重组慢病毒表达载体的构建  

Cloning of ERbeta gene and construction of recombination ERbeta gene lentivirus expression vector

作　　者：李江鹏[1] 赵华栋[1] 张健[2] 李燕[2] 韩腾龙[2] 何显力[1] 

机构地区：[1]第四军医大学唐都医院普外科,陕西西安710038 [2]第四军医大学生物化学与分子生物学实验室,陕西西安710032

出　　处：《现代肿瘤医学》2012年第5期875-878,共4页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(编号:30972930);第四军医大学唐都医院精英人才资助计划项目

摘　　要：目的:构建含人雌激素受体ERbeta基因的重组慢病毒表达载体,为以ERbeta为干预靶点的乳腺癌相关研究奠定基础。方法:经RT-PCR扩增ERbeta基因片段,并将其连接到入门载体pENTRTM3C,然后经LR重组转入到慢病毒表达载体pLenti/V5-DEST中,进而构建含ERbeta基因的慢病毒载体,并进行基因测序鉴定和PCR鉴定。结果:pLenti-ERbeta表达载体测序结果与GenBank中ERbeta基因序列一致,并且其经PCR可扩增出ERbeta基因片段。结论:成功构建携带人ERbeta基因的重组慢病毒表达载体。Objective：To construct the recombinant lentiviral expression vector containing human estrogen receptor ERbeta for research of breast cancer about the ERbeta gene.Methods：Cloned the ERbeta gene by RT-PCR method,and linked it into the pENTRTM3C carrier to create an entry clone,and generate an expression clone by performing an LR recombinantion reaction between the entry clone and a Gateway destination vector（pLenti/V5-DEST）.Finally identified the expression clone（pLenti-ERbeta） by sequencing and PCR.Results：The gene sequence of pLenti-ERbeta expression vector was consistent with that of the ERbeta gene in GenBank database.ERbeta gene fragment of pLenti-ERbeta expression vector could be amplified by PCR.Conclusion： The recombinant of pLenti-ERbeta expression vector could express ERbeta gene and was successfully constructed.

关 键 词：雌激素受体beta 克隆 慢病毒 LR反应 构建 

分 类 号：R730[医药卫生—肿瘤] R737.9[医药卫生—临床医学]