中文摘要：

人MTERF3基因编码线粒体基因转录和能量代谢的负调控因子。采用PCR技术对人MTERF3基因5＇侧翼上游1251 bp启动子序列进行扩增,并将其克隆至荧光素酶表达载体p GL6-TA,构建人MTERF3基因启动子荧光素酶报告基因质粒。经酶切、测序鉴定后,将其用脂质体转染体外培养的HEK293细胞株,利用双荧光素酶测定系统检测其表达活性。研究结果表明,克隆获得的1251 bp DNA序列与Gen Bank报道的一致,且插入方向正确。含人MTERF3基因启动子的报告基因荧光素酶的表达活性显著提高（P〈0.05）,约为对照组（空载体p GL6-TA）的9.8倍。本研究通过对人MTERF3基因启动子的克隆及其荧光素酶表达载体构建与表达活性的测定,为进一步阐明人MTERF3基因表达的调控机制奠定实验基础。

英文摘要：

Human mitochondrial transcription termination factor 3（MTERF3） protein is a negative regulator of human mitochondrial gene expression and energy metabolism.Human MTERF3 gene was amplified with the human genome by PCR,the segment was cloned into the eukaryotic expression vector p GL6-TA.The recombinant was detected by endonuclease and sequenced.Plasmid p GL6-MTERF3-promoter was transfected into HEK293 cells by lipofectamine 2000.The activity of luciferase was detected,and the effect of the human MTERF3 promoter was studied.The sequenced segment（1251bp） in the recombinants was identical to that reported in Gen Bank and the segment was inserted in right direction.The dual-luciferase analysis results showed that the human MTERF3 promoter containing 1251 bp fragment had the high transcriptional activity in HEK293 cells transfected with the recombinant plasmid,which were about 9.8 times of those in negative control group（empty vector p GL6-TA）.The luciferase reporter gene vector with human MTERF3 promoters of 1251 bp has been constructed successfully,which lay a foundation of further study on the regulation of the human MTERF3 gene expression.