利用5′-RACE技术进行人肝再生增强因子相互作用肽cDNA5′末端快速扩增及序列分析

Amplification of 5′cDNA end of human augmenter of liver regeneration by 5′-RACE technique and analysis of sequence

目的:利用5′-RACE技术进行人肝再生增强因子(hALR)相互作用肽cDNA5′末端快速扩增并进行序列分析。方法:按照5′-RACE试剂盒(Takara)操作流程进行hALR相互作用肽cDNA5′末端快速扩增,用BLAST技术对扩增序列进行生物信息学分析。结果:成功获取487bp的5′-RACE反应产物,其中5′端延伸了275bp,同源序列分析表明与人Citron激酶mRNA高度同源。结论:5′-RACE技术可以用于快速有效扩增已知部分cDNA序列的5′末端,Citron激酶可能是与hALR具有相互作用的蛋白。

Objective： To amplify the 5′ cDNA end of human augmenter of liver regeneration （hALR） by 5′ -RACE technique and to analyze the sequence. Methods： 5′ cDNA end of hALR was rapidly amplified by following the instructions of 5′ -RACE kit （Takara）. The acquired sequence was analyzed by bioinformatic tool. Results： 5′ -RACE product with 487bp was successfully obtained, which included extending sequence with 275bp. Homology analysis by BLAST showed highly homologous with human citron kinase. Conclusions： 5′ -RACE method can he used to amplify the 5′ end of partially known cDNA sequence. Citron kinase can interact with hALR.