hR24L基因的克隆及逆转录病毒重组体的构建

Cloning of hR24L gene and the construction of retroviral vector recombinants

目的 克隆人类DNA损伤修复基因hR2 4L ,并构建逆转录病毒表达载体重组体 ,为进一步研究该基因的功能及作用机制奠定基础。方法 用PCR法扩增hR2 4L基因的开放阅读框 (ORF) ,然后连接到 pEGM -T载体中 ,经测序验证无突变后 ,再重组到逆转录病毒表达载体 (pDor -neo)中 ,得到正义和反义重组体。 结果 建立了一种有效的基因克隆方法 ,成功地克隆了hR2 4L基因 ,获得了含有重组体的稳定遗传的细胞株。结论 PCR法是克隆hR2 4L基因的有效方法 。

Objective To clone human DNA repair gene hR24L and to combinate it into the retroviral expression vector pDor.Methods The open reading frame(ORF) of hR24L gene was amplified by PCR,after have been vertified no mutation,then it was cloned into the retroviral expression vector pDor, the recombinant vector was monitor by PAGE.Results hR24L was cloned success and the sense sequence and the anti-sense sequence hR24L-recombinant vector was get.Conclusion This is a efficient method of hR24Lcloning and recombination