大鼠肝癌高表达基因cDNA序列的克隆

Clone of high expression gene cDNA sequence in rats' liver cancer

目的克隆出大鼠肝癌高表达EST片段相关基因的cDNA完整表达序列,为进一步揭示其生物学功能奠定基础。方法通过基因芯片检测出DEN诱发的大鼠肝癌高表达的EST片段;其中部分高表达的EST片段,采用电子克隆与PCR相结合的方法,克隆出其cDNA完整表达序列。结果与正常大鼠肝组织比较,肝癌组织中明显高表达的EST片段有34个。采用EST电子克隆技术与PCR技术相结合的方法,成功克隆出4个EST片段的cDNA序列,经进一步BLAST对比分析,证明G15、G20、G36、G40等4个基因为新基因,并进行GenBank登录,登录号分别为DQ480746、DQ480747、DQ480744、DQ912022。结论利用大鼠基因芯片高通量地筛选出DEN诱发的肝癌组织高表达的有关基因,并在充分利用生物信息学资源的基础上,成功克隆出4个新基因的cDNA序列,为今后进一步深入观察这些基因的功能及其在肝癌发生、发展过程中的作用机制打下良好的研究基础。

Objective To clone the eDNA complete expressive sequence about the gene of the rats liver cancer highly expressing EST fragments,which establishes the foundation to further reveal the biological function. Methods The highly expressive EST fragments of rats＇ liver cancer evoked by DEN by gene chips array were detected,in some of EST fragments, their eDNA complete expressive sequences were cloned by electronic clone and PCR. Results There were 34 obviously highly expressive EST fragments in liver cancer tissue compared with normol liver of rats. The eDNA sequences of 4 EST fragments were triumphantly cloned by electronic clone and PCR. G15,G20,G36 ,G40 were testified as new genes by further contrastive analysis with BLAST and then registered in GenBank whose register numbers were DQ480746, DQ480747, DQ480744, DQg12022,respectively. Conclusions Based on biological information,the research has screened relatively highly expressive genes of liver cancer tissue induced by DEN by gene chips of rats. Then the eDNA sequences of 4 new genes are cloned,which settles good foundations for the further research of function of these genes and their mechanism of action in the pathogenesis and development of liver cancer.