长爪沙鼠ATPase8,ATPase6,COX3基因的克隆及序列分析

Determination and Analysis of ATPase8,ATPase6,COX3 Gene Complete Sequence of Mongolian Gerbil( Meriones unguiculatus)

目的对长爪沙鼠线粒体ATPase8,ATPase6,COX3基因全序列进行测定,并对其进行鉴定及进化分析。方法根据长爪沙鼠已知基因序列设计引物,采用PCR产物测序法,对目的片段进行测序鉴定。结合已公布啮齿类动物ATPase8,ATPase6,COX3基因序列,分析其碱基组成、遗传距离、并基于最小进化法和UPGMA法构建系统进化树。结果获得长爪沙鼠线粒体ATPase8,ATPase6,COX3基因全序列,其与家鼠、小家鼠和仓鼠均具有较高的同源性(76～98%);进化分析结果显示,长爪沙鼠与家鼠、黑家鼠和仓鼠遗传距离较近;碱基G的含量分别为6.9%,10.7%,15.2%,符合mtDNA的特点;A+T含量分别为68.2%,64.1%,59.2%,明显低于G+C含量,符合哺乳动物的特点。结论本研究为首次获得长爪沙鼠ATPase8,ATPase6,COX3基因全序列,长爪沙鼠与家鼠、黑家鼠和仓鼠具有较近遗传距离,本研究为长爪沙鼠进化研究、线粒体的结构和功能研究奠定基础。

Objective To complete sequence of mitochondrial ATPase8,ATPase6,COX3 of Mongolian Gerbil determined basing on the PCR products.The sequence was analysied in phylogenetic tree,nucleotide composition and genetic distance.Methods The primer was designed according to the published sequences.The PCR product were sequenced and determined.Combined with known ATPase8,ATPase6,COX3 sequence of other rodents,nucleotide composition and genetic distance were analysed;phylogenetic tree was constructed by minimum-evolution（ME） methods and UPGMA methods.Result The ATPase8,ATPase6,COX3 gene were cloned in Mongolian Gerbil,which have high homology with rat,mouse and hamster（76~98%）;it is higher genetic relationship with rat,mouse and hamster by phylogenetic analysis;the G content（6.9%,10.7%,15.2%） are similar with feature of mtDNA;the A＋T content（68.2%,64.1%,59.2%） are similar with feature of mammalian genome.Conclusion The ATPase8,ATPase6,COX3,gene sequence of mongolian Gerbil was determined for the first time.We found that Mongolian Gerbil have close genetic relationship with rat,mouse and hamster in this research.This research is an important work for study on evolution,structure and function of the mongolian gerbil mitochondrion.