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人γ-谷氨酰半胱氨酸合成酶催化亚单位基因E-box元件功能初步分析 被引量：2

Preliminary analysis of the E-box element in human γ-glutmylcysteine synthetase catalytic subunit gene
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摘要 目的:探讨人支气管上皮细胞γ-谷氨酰半胱氨酸合成酶催化亚单位(GCLC)基因上游调控序列E-box元件的功能。方法:利用PCR法克隆人GCLC基因上游调控序列,PCR重叠延伸法对2个E-box元件分别和同时进行定点突变。扩增获得的突变体片段克隆入PMD18-T载体,DNA测序鉴定后,亚克隆入野生型GCLC-Luc荧光素酶报道载体。将3个突变体质粒和野生型质粒转染人肺泡上皮细胞A549和人支气管上皮细胞16HBE,检测转染后细胞荧光素酶活性值。结果:测序结果证实,成功将E-box1CACGGG突变为AGCGGG;E-box2CACGTG突变为CACGGA。GCLC-Luc、GCLC-mEbox1-Luc(突变E-box1)、GCLC-mEbox2-Luc(突变E-box2)、GCLC-mEbox12-Luc(突变E-box1和E-box2)转染A549细胞后荧光素酶活性值分别为(5197852±132206)U、(5455692±127431)U、(5315722±244197)U、(5174303±183179)U,转染16HBE细胞后荧光素酶活性值分别为(141157±18907)U、(126454±23724)U、(137315±22179)U、(132441±28970)U。经过统计学分析,各组荧光酶活性没有显著差别,均P>0.05。结论:E-box元件不参与基础状态下A549细胞和16HBE细胞GCLC基因转录调控。
作者 罗伶俐 邹国民 李冰 冉丕鑫
机构地区 广州医学院第一附属医院 广州医学院实验医学研究中心
出处 《中国病理生理杂志》 CAS CSCD 北大核心 2008年第7期1428-1430,共3页 Chinese Journal of Pathophysiology
基金 国家自然科学基金资助项目(No.30170401) 广东省自然科学基金团队基金资助项目(No.05200239)
关键词 16HBE细胞 A549细胞 基因 GCLC Γ-谷氨酰半胱氨酸合成酶 16HBE cells A549 cells Genes,GCLC Gamma-Glutamy1-cysteine synthetase
分类号 R979.1 [医药卫生—药品] R134.4 [医药卫生—劳动卫生]
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参考文献7

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共引文献29

同被引文献19

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中国病理生理杂志
2008年 第7期

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