人甲状腺受体相互作用蛋白15基因全长cDNA的克隆及其特性

Full-length cDNA Cloning and Identification of the Characteristics of Human Thyroid Receptor Interacting Protein 15 Gene

通过生物信息学分析及RT PCR技术 ,从人垂体cDNA文库中克隆到甲状腺素受体相互作用蛋白 15(hTRIP15)的全长cDNA ,长度 1963bp ,编码 4 4 3氨基酸 ,同时克隆该基因不同剪接方式所形成的新的异构体 ,长度 1984bp ,编码 4 50氨基酸 .与基因组序列比较显示该基因具有 12个外显子 ,5号外显子 3′端具有 2个剪切的接点 (-ag) .搜寻UniGene数据库作染色体定位于D15S146 D15S117,该基因在生物进化上具有较高的保守性 ,从单细胞藻类到人类均有该基因同源物表达 ,亚细胞定位为核内 .Northern杂交显示 ,该基因具有 3种不同大小的转录本 ,分别约为 2 0、3 5及 4 0kb ,且在人体各组织中均有一定表达 ,其中骨骼肌、心脏及肾脏组织为高表达 .半定量RT PCR显示在一些内分泌组织均有表达 ,以肾上腺较高 .

By bioinformatics and RT\|PCR procedures,the full\|length cDNA of human thyroid interacting protein 15(hTRIP15),spanning 1 963 bp and encoding 443 amino acids,was cloned from human pituitary cDNA library,and a novel isoform by alternative splicing was also cloned,which had 1 984 bp in length with 450 amino acids.The gene had 12 exons and there were two splicing adaptors(-ag) in the 3′ end of the 5th exon as compared with genomic DNA sequence.hTRIP15 was localized in D15S146\|D15S117 by Unigene searching and conserved in bioevolution from single\|cell organism to human being.The protein encoded by the gene was expressed in nucleus.Northern bloting showed that there were three transcripts in most human tissues and the gene was highly expressed in the skeletal muscle,heart,kidney as well as adrenal tissues.