马黎丽, 陈昌杰, 杨清玲, 章尧. 人端粒酶逆转录酶基因启动子调控表达载体的构建和鉴定[J]. 蚌埠医科大学学报, 2008, 33(6): 665-668.
    引用本文: 马黎丽, 陈昌杰, 杨清玲, 章尧. 人端粒酶逆转录酶基因启动子调控表达载体的构建和鉴定[J]. 蚌埠医科大学学报, 2008, 33(6): 665-668.
    MA Li-li, CHEN Chang-jie, YANG Qing-ling, ZHANG Yao. Construction and identification of a expression vector driven by human telomerase reverse transcriptase gene promotor[J]. Journal of Bengbu Medical University, 2008, 33(6): 665-668.
    Citation: MA Li-li, CHEN Chang-jie, YANG Qing-ling, ZHANG Yao. Construction and identification of a expression vector driven by human telomerase reverse transcriptase gene promotor[J]. Journal of Bengbu Medical University, 2008, 33(6): 665-668.

    人端粒酶逆转录酶基因启动子调控表达载体的构建和鉴定

    Construction and identification of a expression vector driven by human telomerase reverse transcriptase gene promotor

    • 摘要: 目的:构建并鉴定人端粒酶逆转录酶(human telomerase reverse transeriptase,hTERT)基因启动子调控的荧光素酶基因表达载体。方法:用巢式PCR方法克隆hTERT序列长约1100bp的启动子片段,经测序无误后将其插入荧光素酶基因报告载体中,构建pGL3-hTERTp重组质粒,转化JM190细菌得到大量含有hTERT启动子的荧光素酶重组质粒。用双酶切和PCR法鉴定重组质粒,并送测序鉴定。结果:经过酶切和PCR法鉴定成功地构建了携带hTERT启动子的重组荧光素酶基因报告载体。结论:成功获得由hTERT启动子调控的荧光素酶基因表达载体,为研究As2O3对HL-60细胞端粒酶作用的分子机制打下基础。

       

      Abstract: Objective: To construc and identify the expression vector which contains a luciferase gene driven by human telomerase reverse transcriptase(hTERT) gene promotor.Methods: The fragment of hTERT promoter about 1 100 bp was acquired by nested PCR amplification,the correct sequence was inserted into the luciferase gene expression vector.In order to get a lot of containing hTERT promoter luciferase recombinant,the construction of pGL3-hTERTp recombinant was transformed into JM190 bacteria.Restriction endonuclease and PCR were used to identify the recombinant and sequencing.Results: After identifying which used restriction endonuclease and PCR method,we successfully constructed the recombinant of hTERT promoter luciferase gene expression vector.Conclusions: A novel recombinant luciferase gene expression vector driven by hTERT promoter is made successfully,and the newly constructed recombinant is useful for study the molecular mechanism that As2O3 inhibits the expression of telomerase of HL-60 cells.

       

    /

    返回文章
    返回