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目的:克隆并鉴定和分析人Cuedc2启动子,为进一步研究其转录调控机制和功能提供实验基础。方法:对Cuedc2基因翻译起始位点上游约2000bp的序列进行在线生物信息学分析,使用PCR技术扩增该序列并测序,将扩增获得的该片段定向克隆入PGL-3basic载体中,构建荧光素酶报告基因质粒Cuedc2-luc。荧光素酶分析检测启动子的活性。结果:本实验成功构建了含有Cuedc2基因启动子序列的荧光报告系统,经体外验证该报告基因重组载体具有转录活性。结论:本实验所构建的Cuedc2基因启动子报告基因载体,为进一步研究Cuedc2基因的转录调控及其功能奠定了基础。
OBJECTIVE: To clone and identify human Cuedc2 promoter and provide experimental basis for further study of its transcriptional regulation mechanism and function. Methods: Bioinformatics analysis of the sequence about 2000bp upstream of the translation initiation site of Cuedc2 gene was carried out. The sequence was amplified by PCR and sequenced. The amplified fragment was cloned into PGL-3basic vector to construct fluorescence Enzyme reporter plasmid Cuedc2-luc. Luciferase assay detects promoter activity. Results: We successfully constructed a fluorescent reporter system containing the promoter sequence of Cuedc2 gene. The recombinant plasmid was verified to have transcriptional activity in vitro. Conclusion: The Cuedc2 gene promoter reporter gene vector constructed in this study laid the foundation for further study on the transcriptional regulation and function of Cuedc2 gene.