人伴侣蛋白CCTβ亚基全长cDNA的分离与鉴定

Isolation and Identifiction of the Full Length cDNA of the Human Chaperonin CCTβ Subunit

目的 获得人伴侣蛋白CCTβ亚基全长cDNA序列 ,为进一步研究伴侣蛋白在蛋白质折叠中的功能提供理论依据。方法 采用“同源杂交筛选策略” ,运用生物信息学方法结合PCR等分子生物学实验技术 ,从人cD NA文库中分离目的片段 ,并测序鉴定。结果 从人的睾丸组织cDNA文库中分离出一条新的cDNA片段 ,将它与计算机拼接的片段整合后 ,获得一全长为 1935bp的序列 ,该序列含有一个长 16 80bp的开放阅读框 ,编码 5 35个氨基酸 ,并且在其 3′末端有典型的PolyA加尾信号。 结论 基于从该cDNA推导的蛋白质与小鼠的CCTβ亚基氨基酸序列同源度性达 97% 。

Objective To obtain the full length cDNA of human chaperonin CCTβ subunit and provide theoretical basis for the further study of the roles chaperones played in protein folding.Methods Using the homology hybridezation screeing strategy and bioinformatics combined with molecular biology experimental technology such as PCR,the desired sequence was isolated from human cDNA library and sequencing and identification were under-gone.Results A new cDNA sequence was isolated from the cDNA library of the human testis.After integration with the sequence spliced by computer,a full length sequence of 1935 bp was obtained,which contains and open reading frame of 1608 bp,encoding 535 amino acids,with typical polyA end adding signals in its 3' end.Conclusion The deduced protein of the cDNA shares high homology(97%)with the CCTβ subunit from mouse.Therefore the deduced protein encoded by this new gene may be the numan CCTβ subunit.