多功能转录因子CTCF结构域的转录活性研究  

Co-transfection and Transcription Activity Studies of Multifunctional Factor CTCF

作　　者：何莉[1] 朱旭东[1] 黄培堂[1] 

机构地区：[1]军事医学科学院生物工程研究所

出　　处：《生物技术通讯》2006年第5期700-702,共3页Letters in Biotechnology

基　　金：国家科技攻关课题(2004AA002100)

摘　　要：目的:CTCF是广泛存在于真核生物中的多功能转录因子。利用细胞瞬时表达实验在HeLa细胞系中研究CTCF各结构域的转录活性。方法:借用CheckMate哺乳动物双杂交系统的2个质粒pBIND和pG5LUC,将CTCF的N、M、C段及全长编码序列克隆至pBIND质粒的GAL4DNA结合结构域编码序列后的多克隆位点,将表达GAL4DNA结合结构域-CTCF融合蛋白的pBIND重组质粒与由腺病毒晚期主要启动子控制的报告基因质粒pG5LUC共转染HeLa细胞系,检测CTCF各结构域对报告基因表达水平的影响。结果:转染含CTCFN段的质粒可使报告基因的表达量增长至3.5倍以上;转染含CTCF其他片段的质粒对报告基因的表达没有明显的影响。结论:在HeLa细胞中,对启动子具有转录激活活性的主要是CTCF的N段,而M段和C段几乎没有转录激活活性。Objective： CTCF is a multifunctional transcription factor, which is widely expressed in eukaryotes. To know the transcription activities of CTCF domains in HeLa cells. Methods： Two plasmids pBIND and pG5LUC from CheckMate mammalian two-hybrid system were used. The N, M, C fragment or whole CDS of CTCF were inserted into expression plasmid pBIND, which was cotransfected with reporter plasmid pG5LUC into HeLa cells. The expression level of reporter gene luciferase was then detected to indicate the transcription activities of different part of CTCF. Results： pBIND-CTCFN triggered more than 3.5 folds increase of luciferase level relative to the negative control, while other domains of CTCF showed no significant activity. Conclusion： N domain of CTCF has transcription activation function in HeLa cells, while M and C domains have no such an activity.

关 键 词：CTCF 转录因子 细胞瞬时表达实验 转录激活 

分 类 号：Q75[生物学—分子生物学] Q78