[1]吉颖,刘竹,艾青,等.人HAS3基因核心启动子区域的初步鉴定与分析[J].陆军军医大学学报(原第三军医大学学报),2012,34(05):391-395.
 Ji Ying,Liu Zhu,Ai Qing,et al.Preliminary identification and characterization of human hyaluronan synthase 3 core promoter region[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(05):391-395.
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人HAS3基因核心启动子区域的初步鉴定与分析(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
34卷
期数:
2012年第05期
页码:
391-395
栏目:
论著
出版日期:
2012-03-15

文章信息/Info

Title:
Preliminary identification and characterization of human hyaluronan synthase 3 core promoter region
作者:
吉颖刘竹艾青朱江谢濛宇翁华莉刘革力宋方洲卜友泉
重庆医科大学:生物化学与分子生物学教研室,分子医学与肿瘤研究中心,附属第一医院耳鼻喉科
Author(s):
Ji Ying Liu Zhu Ai Qing Zhu Jiang Xie Mengyu Weng Huali Liu Geli Song Fangzhou Bu Youquan
Department of Biochemistry and Molecular Biology, Molecular Medicine and Cancer Research Center, College of Basic Medical Sciences, Department of Otolaryngology, First Affiliated Hospital, Chongqing Medical University, Chongqing, 400016, China
关键词:
透明质酸合成酶3启动子转录调控
Keywords:
hyaluronan synthase 3 promotertranscriptional regulation
分类号:
R394-33;R394.3
文献标志码:
A
摘要:
目的      对人HAS3基因的核心启动子区域进行初步鉴定和分析,为深入研究HAS3基因的转录调控奠定基础。      方法      以前期构建的人HAS3基因启动子荧光素酶报告基因重组体P(-761/-305)为模板,采用PCR介导的基因定点突变技术构建4个不同的系列删除体,并对HAS3启动子区域中的Sp1结合位点和核心启动子元件进行定点突变,构建相应的定点突变重组体;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。      结果        构建了P(-761/-569)、P(-563/-305)、P(-490/-305)和P(-433/-305)4个系列删除体和5个定点突变体;启动子活性分析结果表明,P(-761/-569) 无启动子活性,P(-563/-305)、P(-490/-305)和P(-433/-305)均具有较强的启动子活性,Sp1结合位点和核心启动子元件的定点突变可导致HAS3启动子活性的降低。      结论        HAS3基因的核心启动子区域主要位于其转录起始位点上游附近-433~-305 bp内,Sp1可能在HAS3的转录调控中起重要作用。
Abstract:
Objective        To preliminarily identify and characterize the core promoter region for human hyaluronan synthase 3 (HAS3) gene.        Methods        The previous constructed HAS3 promoter reporter, P(-761/-305), was used as template to make four different deletion mutants and site-directed mutants by using PCR based site-directed mutagenesis. Dual luciferase reporter assay was used to determine promoter reporter activity.        Results        Four deletion mutants were constructed, including P(-761/-569), P(-563/-305), P(-490/-305) and P(-433/-305). Among the four deletion mutants, P(-761/-569) had no promoter activity whereas P(-563/-305), P(-490/-305) and P(-433/-305) showed significant promoter activity. Site-directed mutagenesis of Sp1-binding site and core promoter element resulted in a decrease in HAS3 promoter activity.        Conclusion        The core promoter region of HAS3 gene is mainly located in a 129-bp region (-433 bp to -305 bp) nearby the major transcriptional start site. Sp1 might play an important role in the transcriptional regulation of HAS3 gene.

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更新日期/Last Update: 2012-03-01