小鼠凋亡相关新基因TFAR15的克隆和序列分析

目的克隆人白血病细胞凋亡相关新基因TFAR15的小鼠同源序列 ,并比较其在不同种属间的序列同源性。方法利用EST(expressedsequencetag)拼排、RT PCR、DNA序列测定和计算机分析技术 ,对小鼠TFAR15进行研究。结果首次成功地进行了小鼠TFAR15全长cDNA的克隆和序列分析 ,发现小鼠TFAR15与小鼠其它cDNA没有明显的同源性 ,因此提交GenBank并被收录 ,登录号为AF159368。小鼠TFAR15和人TFAR15在核苷酸水平上有92.3 %的同源性 ,在氨基酸水平上有高达98.6 %的同源性 ;同时发现小鼠TFAR15在氨基酸水平上与线虫C14A4.11蛋白质有38 %的同源性。功能区分析发现 ,小鼠TFAR15cDNA序列含编码212个氨基酸的开放读码框架 ,有2个可能的N 糖基化位点 ,3个可能的caseinkinaseII磷酸化位点 ,4个可能的PKC磷酸化位点 ,并且可能是一种胞浆蛋白(cytoplasmicprotein)。结论小鼠TFAR15是进化上高度保守的新基因 ,可能具有重要的功能。

cDNA cloning and sequencing of a novel mouse apoptosis related gene TFAR15

Aim To clone a mouse homologous sequence of a human novel gene TFAR15(TF 1 cell apoptosis related gene 15) and to analyze their sequence homology among the different species. Methods The EST(expressed sequence tag) sequence alignment method, RT PCR, DNA sequencing, and computer analysis were used for the cDNA cloning of mouse TFAR15. Results The full length of mouse TFAR15 cDNA was successfully cloned and sequenced. There was 92.3%homology between mouse and human TFAR15 at the nucleotide level and 98.6%homology at amino acid level. It contained an open reading frame encoding 212 amino acids, two potential N glycosylation sites, three casein kinase II phosphorylation sites, and four potential PKC phosphorylation sites. It was possible that mouse TFAR15 protein was a cytoplasmic protein. Conclusion Mouse TFAR15 was highly homologous with human TFAR15. Further experiment was needed to confirm its biological activities.