真核细胞转录系统启动T7启动子起始的研究  

Transcriptional initiation of T7 promoter by transcriptional system in eukaryotic cells

作　　者：周天鸿[1] 李月琴[1] 朱嘉明[1] 云泓若[1] 肖小敏[2] 

机构地区：[1]暨南大学生物工程学系,广东广州510632 [2]暨南大学医学院第一附属医院妇产科,广东广州510630

出　　处：《暨南大学学报（自然科学与医学版）》2000年第3期95-99,共5页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学基金!(39770 411)

摘　　要：利用氯霉素乙酰转移酶基因 (CAT)和人低密度脂蛋白受体基因 (LDLR)作为报道基因 ,根据α -鹅膏蕈碱 (α -amanitin)对真核生物RNA聚合酶的选择性抑制 ,分析T7噬菌体启动子为哺乳类动物细胞启动外源基因表达的机制 结果表明 :真核生物RNA聚合酶Ⅱ可启动T7启动子的转录 ;同时应用DNA -蛋白质凝胶泳动技术 ,发现人工合成的T7启动子能与核蛋白质结合 。Using the chlorampheniol acetyltransterase gene and the cDNA of low-density lipoprotein receptor gene as reporters, the function of phage T7 promoter in mammalian cell was studied by inhibition of transcription with α-amanitin, by transcription assay with T7 polymerase in vitro and electrophoretic mobility shift assay(EMSA). The results demonstrated that CAT and LDLR genes as reporters under T7 promoter were transcribed by RNA polymerase Ⅱ. Electrophoretic mobility shift assay revealed that the binding of synthetic T7 promoter with nuclear protein, suggesting further the function of T7 promoter as cis-acting elements of transcriptional system in eucaryotic cells. [

关 键 词：T7启动子 RNA聚合酶Ⅱ 真核细胞 转录系统 

分 类 号：Q754[生物学—分子生物学]