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PNP 表达质粒的构建及鉴定

蔡晓坤, 刘址忠, 林菊生, 马 昕, 梁扩寰

蔡晓坤, 刘址忠, 林菊生, 马 昕, 梁扩寰. PNP 表达质粒的构建及鉴定[J]. 肿瘤防治研究, 2005, 32(07): 414-416. DOI: 10.3971/j.issn.1000-8578.2785
引用本文: 蔡晓坤, 刘址忠, 林菊生, 马 昕, 梁扩寰. PNP 表达质粒的构建及鉴定[J]. 肿瘤防治研究, 2005, 32(07): 414-416. DOI: 10.3971/j.issn.1000-8578.2785
CAI Xiao-kun, LIU Zhi-zhong, LIN Ju-sheng, MA Xin, LIANG Kuo-huan. The Construction and Identif ication of a Vector Containing PNP Suicide Gene[J]. Cancer Research on Prevention and Treatment, 2005, 32(07): 414-416. DOI: 10.3971/j.issn.1000-8578.2785
Citation: CAI Xiao-kun, LIU Zhi-zhong, LIN Ju-sheng, MA Xin, LIANG Kuo-huan. The Construction and Identif ication of a Vector Containing PNP Suicide Gene[J]. Cancer Research on Prevention and Treatment, 2005, 32(07): 414-416. DOI: 10.3971/j.issn.1000-8578.2785

PNP 表达质粒的构建及鉴定

详细信息
    通讯作者:

    林菊生

  • 中图分类号: R73235 + 4

The Construction and Identif ication of a Vector Containing PNP Suicide Gene

  • 摘要: 目的 构建并鉴定一种新型的PNP自杀基因表达载体。方法 将PNP基因插入到pcDNA3.0中,构建PNP基因表达载体pcDNA3.0/PNP;通过酶切、PCR及测序鉴定重组体。采用脂质体介导法将pcDNA3.0/PNP转染三种肿瘤细胞株,RT—PCR检测PNP基因在各细胞株中的表达。结果 目的片段均成功插入相应的载体中,CMV启动子调控下的PNP基因在不同的肿瘤细胞株中实现了表达。结论 CMV启动子调控的PNP基因表达载体是肿瘤基因治疗中一种新型、高效的治疗栽体。

     

    Abstract: Objective  To const ruct and identify a novel expression vector harboring PNP gene. Methods  Inserting PNP gene into pcDNA 3. 0, an expression vector containing PNP gene driven by the CMV promoter, pcDNA 3. 0/ PNP was const ructed by using recombinant DNA techniques. The recombinant was analyzed and identified by restriction enzyme, PCR and sequencing. The expression vector carrying PNP gene was transfected into three tumor cell lines by liposome-mediated method. The expressions of PNP gene in all cell lines were detected by RT-PCR method. Results  The target fragment was cloned into the corresponding vector. The expressions of PNP gene in different tumor cell lines were positive. Conclusion  The expression vector containing PNP gene under the control of a CMV promoter, is an novel effective expression vector for tumor gene therapy.

     

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出版历程
  • 收稿日期:  2004-04-13
  • 修回日期:  2004-09-26
  • 刊出日期:  2005-07-04

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