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Construction of Cox7a2 fluorescent vector and its effect on cytochrome C oxidase activity in mouse Sertoli cell line TM4
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作者 刘保兴 《外科研究与新技术》 2011年第4期260-260,共1页
Objective To construct Cox7a2 fluorescent vector and study its effect on cytochrome C oxidase ( COX) activity in mouse Sertoli cell line TM4. Methods The coding region of CoxTa2 was amplified from mouse Sertoli cell l... Objective To construct Cox7a2 fluorescent vector and study its effect on cytochrome C oxidase ( COX) activity in mouse Sertoli cell line TM4. Methods The coding region of CoxTa2 was amplified from mouse Sertoli cell line TM4 by RT-PCR. PCR product was 展开更多
关键词 line cell Construction of Cox7a2 fluorescent vector and its effect on cytochrome C oxidase activity in mouse sertoli cell line tm4 TM
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Mutated elements of a complex promoter (Amh) can help to demonstrate the role of certain elements in controlling differential gene expression 被引量:1
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作者 David W. Dresser 《American Journal of Molecular Biology》 2012年第4期351-358,共8页
Amh is a single copy gene which is expressed in different ways during mammalian development. Several potential promoter elements have been identified using physiological experimentation and on the basis of interspecif... Amh is a single copy gene which is expressed in different ways during mammalian development. Several potential promoter elements have been identified using physiological experimentation and on the basis of interspecific sequence comparison. The role of putative promoter elements in controlling gene expression has been investigated by many workers over the last two decades and here by individually mutating each element. Expression was measured in vitro in cells of Sertoli descent by flowcytometry using EGFP as a reporter gene. Three lines of murine cells were used;pre- and post-pubertal Sertoli and granulosa cells. Differences between the three lines of cells, support the view that differentiation in this in vitro model system is likely to be at the level of available transcription factors at given points in development. 展开更多
关键词 Anti-Mullerian HORMONE SDM Site Directed Mutation Murine SMAT-1 tm4 sertoli cell lines KK1 GRANULOSA cell line
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Cox7a2荧光载体构建及其对小鼠支持细胞细胞色素C氧化酶活性的影响 被引量:1
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作者 刘保兴 彭胜杰 +3 位作者 刘刚 张圣强 陈亮 王传航 《中华泌尿外科杂志》 CAS CSCD 北大核心 2011年第7期490-493,共4页
目的构建pEYFP-Cl-Cox7a2表达载体,研究其在小鼠睾丸支持细胞(TM4)中的表达及对细胞色素C氧化酶(COX)活性的影响。方法应用RT—PCR方法从TM4细胞克隆Cox7a2,利用BamHI和EcoRI酶切位点将Cox7a2克隆到pEYFP—C1载体,经酶切、测序... 目的构建pEYFP-Cl-Cox7a2表达载体,研究其在小鼠睾丸支持细胞(TM4)中的表达及对细胞色素C氧化酶(COX)活性的影响。方法应用RT—PCR方法从TM4细胞克隆Cox7a2,利用BamHI和EcoRI酶切位点将Cox7a2克隆到pEYFP—C1载体,经酶切、测序及蛋白印迹等技术进行鉴定,并将重组质粒pEYFP—C1-Cox7a2转染TM4细胞后6、12、24、48h,采用分光光度计测定COX活性。结果从TM4细胞克隆到Cox7a2基因完整的编码序列,大小为252bp。构建pEYFP—CI.Cox7a2表达载体,经酶切、测序鉴定证实克隆正确,转染TM4细胞的效率达70%,融合蛋白表达产物相对分子质量为37000。重组质粒转染后6、12、24、48h时COX活性分别为(0.642±0.051)、(0.542±0.049)、(0.311±0.021)和(0.216±0.010)U/mg,不转染组和转染空载体组分别为(0.714±0.064)、(0.653±0.031)U/mg。与不转染组相比,重组质粒转染后12、24、48h组COX活性显著降低(P〈0.05)。结论重组质粒pEYFP—Cl-Cox7a2载体成功构建。CoxTa2基因对TM4细胞COX活性具有抑制作用,可能对调控COX活性发挥重要作用。 展开更多
关键词 电子传递复合物IV COX7A2 pEYFP-C1载体 tm4支持细胞 COX活性
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