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肖永红, 刘殿武, 李青, 陈清轩. 大鼠肝纤维化相关基因的差异表达分析[J]. 中国公共卫生, 2004, 20(7): 791-793.
引用本文: 肖永红, 刘殿武, 李青, 陈清轩. 大鼠肝纤维化相关基因的差异表达分析[J]. 中国公共卫生, 2004, 20(7): 791-793.
XIAO Yong-hong, LIU Dian-wu, LI Qing, . Differential expression analysis on liver-fibrosis-related genes in rats[J]. Chinese Journal of Public Health, 2004, 20(7): 791-793.
Citation: XIAO Yong-hong, LIU Dian-wu, LI Qing, . Differential expression analysis on liver-fibrosis-related genes in rats[J]. Chinese Journal of Public Health, 2004, 20(7): 791-793.

大鼠肝纤维化相关基因的差异表达分析

Differential expression analysis on liver-fibrosis-related genes in rats

  • 摘要:
      目的   寻找大鼠正常肝组织与纤维化肝组织间的差异表达基因, 从基因水平阐明肝纤维化的发生机制。
      方法   采用mRNA差异显示法(mRNA differential display PCR, DD-PCR), 分别提取正常肝组织及纤维化肝组织的总RNA, 进行逆转录PCR, PCR产物于6%变性聚丙烯酰胺凝胶(PAG)上电泳, 放射自显影后寻找两者间的差异片段, 回收差异片段进行2次PCR, 对2次PCR产物采用反向RNA印迹法(Northern杂交)鉴定, 以排除假阳性, 将有差异的PCR产物克隆到质粒载体上, 测序并进行同源性比较。
      结果   从凝胶中共选出16条差异带, 12条获得再扩增。对其中4个差异带进行克隆和反向Northern杂交鉴定, 获得2条有显著性差异的cDNA片段(N2、F1)。同源性分析显示, N2与小家鼠载脂蛋白a5(apoa5)基因序列94%同源, 未发现与F1片段相同的基因序列。
      结论   应用mRNA差异显示技术获得1条与肝纤维化相关的特异表达基因, 对寻找肝纤维化的防治方法具有重要意义。

     

    Abstract:
      Objective   To screen differential expression genes between normal liver and hepatic fibrosis in rats and to demonstrate the molecular mechanism from gene.
      Methods   mRNA differential display PCR (DD-PCR) was used. First both total RNAs were isolated and reversely transcribed into cDNAs respectively. The products of PCR were run on 6% denaturing polyacrylamide gels(PAG)and autoradiographs were examined to find the differential bands between both gels. The differential bands were reamplified using the same PCR condition as before. Reverse northern analysis was used to identify the bands. After sequencing, the DNA sequence was checked with GenBank data for homology.
      Results   16 differential bands were selected from the gel and 12 bands were special by repeated PCR. In the 4 sub-clones, 2 differentially expressed fragments were obtained(N2, F1)by reverse northern hybridization. Homology comparing, the sequence of N2 had 94% homology with Mus musculus apolipoprotein a5(apoa5), but F1 was not found.
      Conclusion   By mRNA differential display, mRNAs from normal liver and hepatic fibrosis were compared to find one differentially expressed liver-fibrosis-related gene fragment. It will be effective to study the gene display to find the special curative method.

     

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