| 人SGK1超表达载体的构建及鉴定 |
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| 引用本文： | 王玉梅,张晓丽,王全胜,冯玉锡. 人SGK1超表达载体的构建及鉴定[J]. 中国中西医结合肾病杂志, 2006, 7(3): 135-137 |
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| 作者姓名： | 王玉梅 张晓丽 王全胜 冯玉锡 |
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| 作者单位： | 华中科技大学同济医学院附属协和医院肾内科,武汉,430022 |
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| 摘 要： | 目的:构建人血清及糖皮质激素诱导型蛋白激酶(SGK1)超表达载体质粒,为SGK1的体外研究提供有效的工具.方法:细胞中RNA经反转录合成cDNA,PCR扩增获取人全长SGK及构建于哺乳动物的表达载体.基因激活型(SD)及灭活型(KN)人SGK1经点突变获取.克隆基因的正确性通过酶切鉴定及DNA测序证实.用磷酸钙沉淀法转染人胚肾293(HEK293)细胞,测定其转染率.结果:构建的人SGK1质粒能在人胚肾细胞中超表达.结论:人SGK1超表达载体的成功构建为糖尿病肾病发病机制的研究提供了新的工具.
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| 关 键 词： | 载体 糖尿病 |
| 收稿时间： | 2005-12-25 |
| 修稿时间： | 2005-12-25 |
Construction and Identification of Over Expressed Human SGK1 |
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| WANG Yumei, ZHANG Xiaoli, WANG Quansheng, et al. Construction and Identification of Over Expressed Human SGK1[J]. Chinese Journal of Integrated Traditional and Western Nephrology, 2006, 7(3): 135-137 |
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| Authors: | WANG Yumei ZHANG Xiaoli WANG Quansheng et al |
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| Abstract: | Objective:To construct plasmid with overexpressed human SGK1 to provide a useful tool for its studies in vitro.Methods:the full length SGK,was amplified with cDNA,cDNA reverse transcriped from cell total-RNA,then constructed into the expression vector.The active and dominant-negative human SGK1 were obtained by site-directed mutagenesis.All SGK1 plasmids were verified by digestion with restriction enzymes and DNA sequencing.The efficiency of transfection of recombinant SGK1 was analysed in HEK293 cells by phosphate-calcium deposit method.Results:Recombinant human SGK1 is successfully over-expressed in the HEK293 cells.Conclusion:The successful construction of over expressed human SGK1 developed a new technique for studying the pathogenesis of diabetes nephrology. |
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| Keywords: | SGK1 |
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