人周围血单核细胞载脂蛋白E基因表达的竞争性逆转录-聚合酶链反应定量分析
DOI:
作者:
作者单位:

作者简介:

通讯作者:

基金项目:

海南省自然科学基金(80084)资助


A Competitive Reverse Transcription-Polymerase Chain Reaction to Study Quantification of Apolipoprotein E Gene Expression in Human Monocyte-derived Macrophages
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
    摘要:

    为建立人周围血单核细胞载脂蛋白E基因表达检测方法,研究载脂蛋白E基因与儿童健康的关系,我们抽取2 6例健康儿童外周静脉血,分离血单核细胞,抽提RNA ,采用逆转录—聚合酶链式反应检测载脂蛋白E基因表达,并以正常人cDNA作定量标准物,待测样品与定量标准物共扩增,计算出待测样本的个体的mRNA量。研究发现载脂蛋白E基因能在健康儿童周围血单核细胞表达,健康儿童载脂蛋白E基因表达量为0 .37±0 .15mol/molmRNA。表明采用竞争性逆转录—聚合酶链反应方法来检测人周围血单核细胞载脂蛋白E基因表达的分析方法快速、简便、灵敏、实用、可靠,且能准确定量,值得推广应用。

    Abstract:

    Aim To study the relationship between the expression status of apolipoprotein E gene and children health,developed a new method for the quantification of apolipoprotein E mRNA in human monocyte-derived macrophages. Methods 10 mL venous blood was obtained from 26 health children.The mononuclear cells were isolated.Total RNA from the monocytes was extracted.Apolipoprotein E gene express were determined by reverse transcription-polymerase chain reaction. After synthesis of the internal apolipoprotein E DNA standard,we amplified internal standard DNA and cDNA of samples studied. Quantification of apolipoprotein E gene express was calculated. Results Apolipoprotein E mRNA was 0.37±0.15 mol/mol GAPDH mRNA in health children. Conclusions The competitive reverse transcription- polymerase chain reaction was a rapid,simple,sensitive method to study expression of apolipoprotein E gene. This method might be a valuable tool to assess apolipoprotein E gene expression in human disease.

    参考文献
    相似文献
    引证文献
引用本文

向伟,马燕琳,符生苗,赵水平,赵迪成,杨进福,陈炽,王福利,王果,聂赛.人周围血单核细胞载脂蛋白E基因表达的竞争性逆转录-聚合酶链反应定量分析[J].中国动脉硬化杂志,2003,11(5):473~475.

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
历史
  • 收稿日期:2002-11-27
  • 最后修改日期:2003-06-19
  • 录用日期:
  • 在线发布日期: