携带SKP2基因的shRNA表达质粒的构建与鉴定
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宋茂民

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Construction and appraise of SKP2 gene shRNA expression plasmid
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    摘要:

    目的:构建含人SKP2基因不同位点的一系列shRNA真核表达质粒,并进行酶切鉴定、测序。
    方法:设计合成3对SKP2基因干扰寡核苷酸序列,形成双链后将其依次连入带有U6启动子的pSIREN-RetroQ载体,构建成能产生SKP2短发卡RNA的质粒。
    结果:构建的核苷酸序列经鉴定构建成功,能成功的克隆入带有pSIKEN RetroQ的载体中。
    结论:成功构建并鉴定了针对人SKP2基因3个不同位点的shRNA的真核表达质粒,为SKP2为靶点的肿瘤基因治疗研究奠定了基础。

    Abstract:

    Abstract:Objective:To construct and appraise a series of shRNA eukaryotic expression plasmid which contains vary spots of human SKP2 gene.
    Methods :Three pairs of hairpin-like oligonucleotide sequences specific for human SKP2 gene were designed and synthesized. They were cloned into the eukaryotic expression pSIREN-RetroQ plasmid. The recombinated plasmids were confirmed by PCR and sequencing.
    Results:The shRNA sequences were constructed successfully, and could be inserted into the eukaryotic expression vector pSIREN-RetroQ.
    Conclusions:Three shRNA transcripting plasmids pSIREN-RetroQ are successfully constructed, and form a basis for research in cancer gene target therapy against SKP2 gene.

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任金帅, 程石, 宋茂民.携带SKP2基因的shRNA表达质粒的构建与鉴定[J].中国普通外科杂志,2008,17(3):12-252.
DOI:10.7659/j. issn.1005-6947.2008.03.012

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  • 在线发布日期: 2008-03-25