生命科学研究2017，Vol.21Issue(4)：289-294,6.DOI:10.16605/j.cnki.1007-7847.2017.04.002
肝癌细胞hMTERF3基因启动子表达载体的构建及其受DNA甲基化的影响
Construction of a Reporter Vector Regulated by hMTERF3 Promoter and Effect of DNA Methylation on hMTERF3 Promoter Activity in Hepatoma Cells
摘要
Abstract
To construct a reporter vector regulated by the hMTERF3 promoter to determine the effect of DNA methylation on the promoter activity of hMTERF3 gene,PCR amplification was performed to obtain seven potential hMTERF3 promoter fragments,which were then cloned into the vector to obtain recombinant constructs.Promoter activities of different hMTERF3 fragments in cancer cells were detected by luciferase assay to identify the potential promoter area.The activity of the hMTERF3 promoter with or without treatment of methylase M.Sss Ⅰ was also evaluated by luciferase assay.It showed that three fragments (P523,P866 and P932),all of which contained the P523 sequence,had stronger promoter activity than the pGL6 control.Their promoter activities decreased significantly after methylase treatment (P＜0.05).A luciferase reporter gene system containing the hMTERF3 promoter was successfully constructed.P523 is the potential core area of the hMTERF3 promoter.These results provide a basis for studying the epigenetic mechanism of hMTERF3 gene expression in hepatoma cells.关键词
肝癌细胞/线粒体转录终止因子3基因(hMTERF3)/启动子/载体构建/甲基化Key words
hepatoma cells/human mitochondrial transcription termination factor 3 (hMTERF3)/promoter/plasmid construction/methylation分类
生物科学引用本文复制引用
熊伟,孙美涛,梅雯,自加吉,杨勇琴,左绍远,张晓娟..肝癌细胞hMTERF3基因启动子表达载体的构建及其受DNA甲基化的影响[J].生命科学研究,2017,21(4):289-294,6.基金项目
国家自然科学基金资助项目(81560458,31601155) （81560458,31601155）
云南省教育厅科学研究基金重点资助项目(2014Z126) （2014Z126）
云南省中青年学术和技术带头人后备人才资助项目(No.19) （No.19）
大理大学大学生创新创业计划资助项目(CXCY-X-2016-13) （CXCY-X-2016-13）
