大绒鼠解偶联蛋白1基因部分序列扩增与分析

解偶联蛋白1（Uncoupling protein 1,UCP1）是位于褐色脂肪组织线粒体内膜上的一种解偶联蛋白,该蛋白可以诱导质子漏从而产热。通过设计简并引物进行RT-PCR从大绒鼠BAT中获得UCP1基因cDNA核心序列,RTPCR所得产物长约458 bp,包含的开放阅读框（open reading frame,ORF）为456 bp,编码151个氨基酸。通过BLAST搜索,所得大绒鼠UCP1基因cDNA氨基酸序列与黑线仓鼠、橙腹草原田鼠、金黄仓鼠、小家鼠和褐家鼠等哺乳动物的UCP1氨基酸序列同源性均在80%以上,而与鱼类和两栖类的氨基酸序列同源性在61%以下。研究结果表明UCP1在哺乳类中高度保守。同时,通过NJ方法以UCP1序列构建系统进化树表明大绒鼠与橙腹草原田鼠聚成一支,构成田鼠类分支。

Amplification and analysis of partial sequence of UCP1 in Eothenomys miletus

Uncoupling protein 1（ UCP1） is located in the inner mitochondrial membrane of brown adipose tissue（ BAT）,it can dissipate the proton motive force across the inner mitochondrial membrane as heat. Partial cDNA sequences of UCP1 from BAT in Eothenomys miletus was obtained using the RT-PCR method. The obtained partial sequences of UCP1 was 458 bp in length containing open reading frames（ ORF） of 456 bp,151 amino acids. A search of the BLAST database showed that the deduced sequence of UCP1 in E.miletus was very similar to that of Cricetulus griseus,Microtus ochrogaster,Mesocricetus auratus,Mus musculus,Rattus norvegicus and other mammals,and alignment of the deduce amino acid sequences was more than 80%. However,alignment of the deduced amino acid sequence with fish and amphibians was less than 61%. These results suggested that UCP1 genes were conserved highly in mammals. Moreover,it showed that E. miletus had a close relationship with M. ochrogaster based on the UCP1 gene which were used to reconstruct phylogenetic trees by using NJ method,and they constituted a branch of Microtus.