Site-directed mutagenesis of cis-acting element, CHR, in human NFBD1 promoter region

朱江; 兰欢; 洪苏玲; 卜友泉

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Site-directed mutagenesis of cis-acting element, CHR, in human NFBD1 promoter region

更新时间：2026-01-09

- Site-directed mutagenesis of cis-acting element, CHR, in human NFBD1 promoter region
- Journal of Army Medical University Vol. 32, Issue 10, Pages: 1020-1023(2010)
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  CLC： R394-33;R394.3
- Published：2010
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朱江,兰欢,洪苏玲,卜友泉.人NFBD1启动子区域顺式作用元件CHR的定点突变分析[J].陆军军医大学学报(原第三军医大学学报),2010,32(10):1020-1023.

　Zhu Jiang,Lan Huan,Hong Suling,Bu Youquan.Site-directed mutagenesis of cis-acting element, CHR, in human NFBD1 promoter region[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(10):1020-1023.
朱江,兰欢,洪苏玲,卜友泉.人NFBD1启动子区域顺式作用元件CHR的定点突变分析[J].陆军军医大学学报(原第三军医大学学报),2010,32(10):1020-1023. DOI：

　Zhu Jiang,Lan Huan,Hong Suling,Bu Youquan.Site-directed mutagenesis of cis-acting element, CHR, in human NFBD1 promoter region[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(10):1020-1023. DOI：

摘要

目的对NFBD1启动子区域顺式作用元件CHR进行定点突变分析，以分析CHR在NFBD1转录调控中的作用。方法以原有NFBD1核心启动子荧光素酶报告基因重组体NFBD1-PS1-325为模板，采用PCR介导的基因定点突变技术对NFBD1启动子区域中的CHR位点进行定点突变，构建相应的CHR定点突变重组体；采用荧光素酶双报告基因分析技术检测各重组体的启动子活性，分析CHR定点突变对NFBD1启动子活性的影响；结合阿霉素处理，分析CHR在DNA损伤后NFBD1转录下调中的潜在作用。结果 CHR定点突变后能显著降低NFBD1的启动子活性；CHR定点突变后显著减弱了DNA损伤后NFBD1的转录下调比例。结论 CHR参与NFBD1的基础转录调控及DNA损伤后的转录下调。

Abstract

Objective To investigate the role of a cis-acting element

cell cycle genes homology region (CHR)

in the transcriptional regulation of human NFBD1 gene (a nuclear factor with BRCT domain 1) by conducting a site-directed mutagenesis analysis on the element in human NFBD1 promoter region. Methods Wild type of NFBD1 promoter reporter

NFBD1-PS1-325

was used as template to make CHR mutant by using PCR based site-directed mutagenesis. Dual luciferase reporter assay was used to determine promoter reporter activity. Adiamycin treatment was employed to investigate the role of CHR in the transcriptional downregulation of NFBD1 after DNA damage. Results Site-directed mutagenesis of CHR caused a significant decrease in NFBD1 promoter activity

and also attenuated the transcriptional down-regulation of NFBD1 after DNA damage. Conclusion CHR element might be involved in both basic transcriptional regulation and transcriptional downregulation of NFBD1 after DNA damage.

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