Caveolin-1基因重组腺病毒表达系统的构建与鉴定  

Construction and identification of adenovirus expressing caveolin-1

作　　者：李二毛 沈飞[2] 张焕玲[2] 蔡文松[2] 朱光辉[2] 刘启才[1] 徐波[2] 

机构地区：[1]广州医科大学实验医学中心,广东广州510182 [2]广州市第一人民医院普通外科,广东广州510182

出　　处：《广州医科大学学报》2014年第4期9-12,18,共5页Academic Journal of Guangzhou Medical University

基　　金：广州市医药卫生科技重点项目(201102A212018);王宝恩肝纤维化研究基金(20100020);广州市卫生局中西医结合科研项目(20112A011010);广东省科技计划项目(2009B060700019)

摘　　要：目的:构建靶向肝脏的小凹蛋白-1 caveolin-1基因重组腺病毒并对该表达体系进行鉴定。方法:根据GeneBanks中caveolin-1的序列设计引物,以大鼠肝组织提取总RNA为模板,PCR扩增caveolin-1,并将caveolin-1序列克隆至pAdTrack-CMV质粒中,再与pAd-easy质粒进行同源重组构建重组腺病毒载体,利用脂质体法于293细胞中进行包装重组腺病毒,RT-PCR检测caveolin-1基因的转录,Westernblotting检测caveolin-1蛋白的表达情况。结果:PCR及Western blotting检测结果表明成功构建出caveolin-1基因重组腺病毒。结论:成功构建出caveolin-1基因重组腺病毒,为进一步研究caveolin-1基因在肝纤维化及肝硬化疾病中的作用机制打下实验基础。Objective：To construct and identify the target liver adenovirus expressing caveolin-1. Methods：According to Gene Banks Sequence, the primers of caveolin-1 sequence were designed, genomic RNA from rat liver tissue was extracted as a template, caveolin-1 was amplified by PCR, and the sequence of caveolin-1 was cloned into pAd Track-CMV plasmid, and homogonously recombined with pAd-easy plasmid to obtain the recombinant adenoviral vector. Recombination adenovirus was produced in 293 cells by lipofectamine method, transcription of caveolin-1 and expression of caveolin-1 were detected by RT-PCR and Western blotting, respectively. Results： Detection results of PCR and Western blotting showed that expressing caveolin-1 recombination adenovirus were prosperity constructed. Conclusion： Successfully constructed caveolin-1 recombination adenovirus helps further study of the effects of caveolin-1 in liver fibrosis and liver cirrhosis.

关 键 词：小凹蛋白-1 腺病毒 肝纤维化 大鼠肝组织 CAVEOLIN-1 

分 类 号：R363[医药卫生—病理学]