HPV16 E6基因特异性RNA干扰表达载体的构建及对HPV16 E6基因抑制作用的研究
DOI:
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家自然科学基金资助(30371483)


Construction of HPV16-E6 specific RNA interference expression vector and study of its inhibitory effect
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
  • |
  • 文章评论
    摘要:

    目的:构建针对HPV16 E6基因的RNA干扰表达载体,并研究其对宫颈癌HPV16 E6基因的抑制作用-方法:针对HPV16 E6基因序列设计shRNA(short hairpin RNAs,shRNAs)片断,构建针对HPV16 E6基因的RNA干扰(RNA interference, RNAi)质粒表达载体,脂质体法转染宫颈癌Caski细胞株,应用荧光定量PCR及流式细胞术检测其对HPV16 E6 mRNA及蛋白表达的影响-结果:经PCR及测序证实3种表达载体构建成功,细胞试验表明3种表达载体均抑制了HPV16 E6的mRNA及蛋白的表达,其中CaskiB细胞HPV16E6mRNA的抑制率为89.5%,蛋白抑制率达98.1%-结论:RNAi表达载体可以有效的抑制HPV16 E6基因的表达-

    Abstract:

    Objective:To obtain recombinant HPV16 E6 specific RNA interference expressi-on vectors and investigate the inhibitory effects of the vectors on the expression of human papil-omavirus E6 gene. Methods:According to the computer aided design,56nt oligonucleotide fragments containing different HPV16 E6 specific sequences were synthsized and cloned into the expression vectors,The recombinants were transfected into cervical cancer cell line,Caski,with liposomes. Expression of E6 was detected by FQ-PCR(fluorescence quantitative PCR) and flow cytometry. Results:HPV16 E6 specific siRNA expression vectors were confirmed by PCR analysis and DNA sequencing. Three kinds of expression vectors could reduce the expressions of E6 mRNA and protein all in Caski-B cell. The inhibition ratio of E6 mRNA reduced to 89.5%, and protein inhibition ratio reached 98.1%. Conclusion:The RNAi expression vectors can effectively inhibit the expression of HPVE6 Gene.

    参考文献
    相似文献
    引证文献
引用本文

李大可,彭芝兰,尤志学. HPV16 E6基因特异性RNA干扰表达载体的构建及对HPV16 E6基因抑制作用的研究[J].南京医科大学学报(自然科学版),2008,28(11):1393-1397

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期:
  • 出版日期: