文章摘要
徐瑞瑞,李毓雯,胡平平,田安然,李 军,朱传龙.人EFTUD2启动子的鉴定及初步分析[J].南京医科大学学报,2020,(4):489~495
人EFTUD2启动子的鉴定及初步分析
Structural and functional analysis of the human EFTUD2 promoter
投稿时间:2019-07-08  
DOI:10.7655/NYDXBNS20200406
中文关键词: EFTUD2  剪接体  启动子
英文关键词: EFTUD2  spliceosome  promoter
基金项目:国家自然科学基金(81770591,81800778);江苏省医学重点人才(ZDRCA2016007)
作者单位
徐瑞瑞 南京医科大学第一附属医院感染病科江苏 南京 210029 
李毓雯 南京医科大学第一附属医院儿科江苏 南京 210029 
胡平平 南京医科大学第一附属医院感染病科江苏 南京 210029 
田安然 南京医科大学第一附属医院感染病科江苏 南京 210029 
李 军 南京医科大学第一附属医院感染病科江苏 南京 210029 
朱传龙 南京医科大学第一附属医院感染病科江苏 南京 210029 
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中文摘要:
      目的:鉴定并分析EFTUD2(elongation factor Tu GTP?binding domain?containing 2)基因的启动子区域。方法:应用生物信息学技术对EFTUD2基因结构及潜在启动子区域进行分析,预测4个EFTUD2启动子序列。使用全基因合成技术以及高保真PCR扩增得到目的启动子序列,通过BglⅡ和NheⅠ双酶切,将目的条带克隆到双荧光素酶报告基因载体psiCHECK?2中,得到4个长度不同并覆盖EFTUD2基因转录起始位点附近约2 kb的EFTUD2基因启动子双荧光素酶报告基因重组体。荧光素酶分析检测启动子活性。结果:经酶切、测序鉴定,成功构建了人EFTUD2启动子荧光素酶报告基因重组质粒。通过荧光素酶活性检测,重组体EFTUD2?0.5的相对荧光素酶活性增加(P < 0.05),提示EFTUD2基因核心启动子序列位于转录起始位点附近约500 bp的区域内。结论:EFTUD2基因转录起始位点上游500 bp具有启动子活性,初步判定其核心启动子位于该区域。
英文摘要:
      Objective:To identify and analyze the elongation factor Tu GTP?binding domain?containing 2(EFTUD2)promoter region for the further study of the gene in transcriptional control and expression. Methods:The EFTUD2 gene structure and potential promoter regions were analyzed by bioinformatics methods,and based on which four EFTUD2 promoter sequences were predicted. Whole?genome synthesis technology and high?fidelity PCR amplification were used to obtain the target promoter sequence. The target band was cloned into the dual luciferase reporter vector psiCHECK?2 by BglⅡ and NheⅠ double digestion,and four different EFTUD2 gene promoter luciferase reporter recombinants covering about 2 kb upstream of the EFTUD2 gene transcription initiation site were obtained. Luciferase assay was used to detect promoter activity. Results:Luciferase reporter assays demonstrated that,compared with the control group,the relative luciferase activity of the EFTUD2?0.5 recombinant increased(P < 0.05),suggesting that the EFTUD2 core promoter may be located in a region of 500 bp near the transcription initiation site. Conclusion:It was identified that the 500 bp upstream of the EFTUD2 gene transcription initiation site has promoter activity,and it is speculated that its core promoter is located in this region.
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