安徽医科大学学报2016，Vol.51Issue(5)：620-623,624,5.
大鼠ASIC1基因启动子荧光素酶报告质粒的构建及其功能鉴定
Construction and identification of the rat ASIC1 promoter luciferase reporter plasmid
摘要
Abstract
gene plasmid, and then identify its function. Methods Primers were designed and synthetised, ASIC1 promoter fragment from rat genome DNA was replicated by polymerase chain reaction ( PCR ) . The luciferase report gene pGL3-Basic reporter vector and ASIC1 promoter were digested with restriction enzymes NheI and XhoI separately, and then ASIC1 promoter was connected to pGL3-Basic reporter vector. 293T cells were transiently co-transfected with the constructed pGL3-ASIC1-promoter plasmid and pRL-TK control plasmid, and then detected for luciferase activity after 48 hours. Results Rat ASIC1 gene promoter was amplified by PCR and pGL3-ASIC1-promoter re-porter vector was successfully constructed, and the result of colony PCR and sequencing analysis of recombined plasmid were correct. The transcriptional activity of pGL3-ASIC1-promoter plasmid group was significantly in-creased compared to that of pGL3-Basic plasmid group ( P<0 . 01 ) . Conclusion The rat ASIC1 promoter lucifer-ase reporter gene vector can be successfully constructed, which provides a pivotal basis for further study of regulato-ry mechanism of ASIC1 gene in transcription.关键词
ASIC1/启动子/荧光素酶/报告基因Key words
ASIC1/promoter/luciferase/reporter gene分类
医药卫生引用本文复制引用
周仁鹏,吴小山,王志森,谢亚亚,李悦,代贝贝,王志强,葛金芳,陈飞虎..大鼠ASIC1基因启动子荧光素酶报告质粒的构建及其功能鉴定[J].安徽医科大学学报,2016,51(5):620-623,624,5.基金项目
国家自然科学基金(编号:81271949) （编号:81271949）
