| 刘淑娟1 王涛2 刘朵朵1 乔谷媛1 韩军涛3△.人Cuedc2启动子区的克隆与鉴定*[J].,2014,14(20):3828-3830 |
| 人Cuedc2启动子区的克隆与鉴定* |
| Identification and Analysis of Human Cuedc2 Promoter* |
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| DOI： |
| 中文关键词: Cuedc2 荧光素酶报告系统 转录调控 |
| 英文关键词: Cuedc2 Promoter reporter Transcriptional regulation |
| 基金项目:陕西省科技攻关基金项目（2011K12-10） |
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| 中文摘要: |
| 摘要目的：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。 |
| 英文摘要: |
| ABSTRACT Objective:To clone, identify and analyze human Cuedc2 promoter region and provide experimental basis for the
research on the regulatory mechanism and function ofCuedc2 . Methods:Appoximately 2000 bp up from the ATG of gene Cuedc2 was
bioinfomatics analyzed online. The fragment was generated by polymerase chain reaction and sequenced for correctness. The Cuedc2-luc
promoter reporter was constructed by directly cloned this fragment into PGL-3basic vector and the promoter activity was measured by
luciferase assay. Results:The fragment of 1982bp in upstream of the ATG, was amplified successfully and cloned directly into
PGL-3basic vector to make Cuedc2 promoter reporter. Luciferase reporter assay showed that the fragment of 1956bp in length indeed had
strong promoter activity.Conclusion: The luciferase reporter system of Cuedc2 gene promoter was constructed successfully and could be
used for further study of the function and transcriptional regulation of Cuedc2. |
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