小鼠CSE基因启动子克隆及其活性测定  

Cloning and activity of mouse cystathionine γ-lyase gene promoter

作　　者：王茂先[1] 

机构地区：[1]韩山师范学院生物学系,广东潮州521041

出　　处：《基础医学与临床》2014年第9期1184-1188,共5页Basic and Clinical Medicine

基　　金：国家重点基础研究发展计划(973)项目(2010CB912604);韩山师范学院博士启动项目(QD20140102)

摘　　要：目的研究克隆了小鼠CSE基因启动子,并分析和检测了其调控活性。方法以小鼠血液基因组DNA为模板,克隆了小鼠CSE基因启动子序列,回收纯化,直接连接p GL4.12载体,测序。利用生物信息学软件,分析了CSE基因启动子序列的正确性,利用瞬时转染的方法测定报告基因的活性,根据荧光强度的相对值的大小测定启动子活性。结果发现CSE启动子上转录因子结合位点转录因子结合位点92分以上的有25个。其中,GATA有4个,SRY有3个,USF有2个,MZF1有2个,v-Myb有2个,Cdx A有2个,TATA有1个,AML-1a有1个,RORalp有1个,C/EBP有1个,Nkx-2有1个,Lyf-1有1个,N-Myc有1个,HSF2有1个,E2F有1个。CSE基因调控区没有发现Cp G岛。结论研究结果为进一步CSE基因转录和表达调控机制的研究奠定了实验和理论基础。Objective The promoter sequences of mouse CSE gene were cloned and its transcriptional regulatory mechanisms were studied. Methods The promoter sequence of CSE gene was cloned from the mouse blood genomic DNA by PCR, and was jointed into the pGL4. 12 cloning vector after purification. The sequences of CSE gene promoter were analyzed by a series of bioinformatics software, and the reporter gene activity was tested transient transfection, and promoter activity has been measured and compared to the blank reporter vector in both cell lines. Results There were 25 transcription factor binding sites with more than 92 scores （four GATAs, three SRYs, two USFs, two MZFls, two v-Mybs, two CdxAs, one TATA, one AML-1 a, one RORalp, one C/EBP, one Nkx-2, one Lyf-1, one N-Myc, one HSF2, and one E2F） by analyzing regulator sequence of CSE gene promoter but CpG island was not detected in CSE regulator region. Conclusions The results provide basis for further study of the transcriptional and expressional regulation mechanism the experimental and theoretical of the CSE gene.

关 键 词：小鼠CSE基因 启动子 基因序列分析 转录活性分析 

分 类 号：Q78[生物学—分子生物学]