人MTERF4基因cDNA的克隆及编码蛋白质的生物信息学分析

Cloning and bioinformatic analysis of mitochondrial transcription termination factor 4(MTERF4) cDNA in Homo sapiens

[目的]克隆人线粒体转录终止因子4（mitochondrial transcription termination factor 4,MTERF4）基因cDNA编码区,预测和分析其编码蛋白质的结构与功能。[方法]采用RT-PCR技术扩增人MTERF4基因cDNA的编码序列,采用生物信息学方法对人MTERF4蛋白进行分析。[结果]人MTERF4基因cDNA编码序列为1 146bp,编码一个由381氨基酸组成的亲水性蛋白质,分子量为45.78 k D,理论等电点为9.49。该蛋白N端1-42个氨基酸为前导肽序列,定位于细胞线粒体。其二级结构主要为α螺旋和无规则卷曲,包含4个MTERF基序。三级结构预测结果与二级结构相符。蛋白质多重序列比对和聚类分析显示,人MTERF4蛋白与大鼠、小鼠的MTERF4蛋白具有高度同源性,在系统发育树上聚为一类。[结论]人MTERF4基因cDNA的克隆及编码蛋白质的生物信息学分析为进一步研究其生物学功能奠定了理论依据。

[Objective]Human mitochondrial transcription termination factor 4（MTERF4） gene was cloned and analyzed the structure and functional characteristics of human MTERF4 by bioinformatics method. [Methods]RT-PCR was used to cloned the cDNA coding sequence of human MTERF4,many bioinformatic methods were used to analysis its structure and function. [Results]The results showed that,the ORF of human MTERF4 cDNA was 1 146 bp,encode the hydrophilic protein consist of 381 amino acids. The calculated molecular mass of human MTERF4 was 45. 78 k D. The theoretical isoelectric point was 9. 49. Human MTERF4 localized in mitochodria,and the N-termination 1-42 aa were transit peptide. Main composition of the protein secondary structure were（-helix and random coil,it contains four MTERF motifs. The multiple alignment and clustering analysis showed that,human MTERF4 is a orthologous protein belong to MTERF family,and have high similarity to Rattus norvegicus and Mus musculus MTERF4. [Conclusion]The cloning of human MTERF4 and bioinformatic analysis results provide bioinformation which will helpful for the further study of the biological function of h MTERF4.