雌雄驯鹿茸皮组织ENO1基因cDNA克隆与表达  

cDNA Cloning and Expression of ENO1 Gene in Antler Tip Skin between Female and Male Reindeer

作　　者：韩若冰 韩磊 王强辉 李和平[1] Han Ruobing;Han Lei;Wang Qianghui;Li Heping(Northeast Forestry University,Harbin 150040,P.R.China)

机构地区：[1]东北林业大学,哈尔滨150040

出　　处：《东北林业大学学报》2019年第1期92-95,100,共5页Journal of Northeast Forestry University

基　　金：国家林业局珍稀濒危物种野外救护与繁育项目(2012);中央高校基本科研业务专项资金项目(2572016AA42)

摘　　要：选用雌、雄驯鹿(Rangifer tarandus)顶端茸皮组织为试验材料,参考Genebank数据库中白尾鹿(Odocoileus virginianus)、牛(Bos taurus)、羊(Ovis aries)等同源物种的ENO1基因序列设计同源引物,采用PCR及T-A克隆技术以雄性驯鹿茸皮组织为材料克隆ENO1基因的c DNA序列,并运用荧光定量PCR技术对ENO1基因在雌、雄驯鹿茸皮组织中的表达量进行对比分析。结果表明:成功克隆获得驯鹿ENO1基因,其编码区片段大小为1 305 bp,共编码434个氨基酸;ENO1基因编码的蛋白的相对分子质量为47 342.09,其二级结构以α-螺旋为主;Blastp比对显示,驯鹿ENO1基因与羊、白尾鹿、牛的同源性最高,均为99%。构建系统进化树发现,驯鹿ENO1基因与白尾鹿的亲缘关系最近,与白头鹰(Haliaeetus leucocephalus)的亲缘关系最远;荧光定量PCR结果显示,ENO1基因在茸皮组织中的表达量在雌、雄之间存在差异,其中在雄性驯鹿茸皮组织中的表达量是雌性驯鹿茸皮组织中的(3.35±0.12)倍。With the antler tip skin of female and male reindeer(Rangifer tarandus),the homologous primers were designed based on the ENO1 gene sequence of white-tailed deer(Odocoileus virginianus),cattle(Bos taurus),and sheep(Ovis aries)in GeneBank database.PCR and T-A cloning were used to obtain the cDNA sequence of ENO1 gene in the antler tip skin of male reindeer,and fluorescent quantitative PCR was used to compare the expression level of ENO1 gene in antler tip skin between female and male reindeer.The size of coding area fragment was 1 305 bp and encoded a total of 434 amino acids.The ENO1 protein was a hydrophilic protein of 47 342.09 and the secondary structure is dominated byα-helix.The homology of the reindeer ENO1 gene to sheep,white-tailed deer,and cattle was up to 99%by the Blastp comparison.Construction of phylogenetic tree revealed that the relationship between ENO1 gene and white-tailed deer is the closest,and the genetic relationship with the white-headed sea eagle(Haliaeetus leucocephalus)is the farthest.By the quantitative real-time PCR,the expression of ENO1 gene in antler tip skin was different between male and female,and the expression level in the antler tip skin of male reindeer was 3.35±0.120 times higher than that in female.

关 键 词：驯鹿 ENO1基因 荧光定量PCR 

分 类 号：S825[农业科学—畜牧学] Q785[农业科学—畜牧兽医]