采用报告基因检测HSV-1潜伏相关启动子在神经细胞的特异性激活  

Detection of Specific Activation of HSV-1 Latency Associated Promoter in Neuronal Cells by Reporter Assays

作　　者：郭景霞[1] 李永鑫[1] 朱慧[1] 史茜[1] 张彩荣[1] 朱红娟[1] 马正海[1] 

机构地区：[1]新疆大学生命科学与技术学院,新疆生物资源基因工程重点实验室,乌鲁木齐830046

出　　处：《中国生物化学与分子生物学报》2011年第9期873-878,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：新疆维吾尔自治区高校科研计划重点项目(No.XJEDU1010I11);新疆生物资源基因工程重点实验室开放课题;新疆大学博士科研启动基金~~

摘　　要：1型单纯疱疹病毒(herpes simplex virus type 1,HSV-1)潜伏相关转录本启动子(latencyassociated transcripts promoter,LATP)在病毒于神经组织中潜伏感染期间保持活性.本研究利用PCR技术扩增获得HSV-1的LATP,并将LATP分别插入增强型绿色荧光蛋白(enhanced greenfluorescent protein,EGFP)基因和荧光素酶基因前,获得真核表达质粒pcDNA/LATP/EGFP和pGL3/LATP/Luc.pcDNA/LATP/EGFP转染293T细胞后,检测到绿色荧光蛋白基因的表达,表明LATP具有启动子活性,其活性低于PCMV.继而以pcDNA/LATP/EGFP和pGL3/LATP/Luc分别转染HeLa细胞、Eca109细胞、U251细胞和SK-N-SH细胞.结果表明,绿色荧光蛋白基因和荧光素酶基因在U251和SK-N-SH两种神经细胞中表达水平极显著(P<0.01),高于其在HeLa和Eca109两种非神经细胞中的表达.说明LATP能够驱动基因在神经组织细胞中高效表达,可作为神经组织特异性表达元件.The herpes simplex virus type 1（HSV-1）maintain the activity of latency associated transcripts promoter（LATP） during latent infection of neurons.In this study,LATP was cloned by PCR and inserted in front of the enhanced green fluorescent protein（EGFP） gene and luciferase gene,respectively.The recombinant plasmid pcDNA/LATP/EGFP and pGL3/LATP/Luc were transfected into 293T cells to measure promoter activities.The results demonstrated that the LATP activity was lower than that of PCMV.We also transfected pcDNA/LATP/EGFP or pGL3/LATP/ Luc into HeLa,Eca109,U251 and SK-N-SH cells,and found that the expression levels of EGFP and luciferase in the SK-N-SH cells and U251 cells were significant higher than those in the HeLa cells and Eca109 cells（P0.01）.These results suggest that LATP could drive gene expression more efficiently in neuronal cell,and might act as an element of neuronal-specific gene expression.

关 键 词：1型单纯疱疹病毒 潜伏相关转录本 启动子 神经组织 

分 类 号：Q936[生物学—微生物学]