东北狍PTN基因cDNA克隆及表达分析  

cDNA Cloning and Expression Analysis of PTN Gene from Northeast Roe Deer

作　　者：卢斌山 王强辉 张强 高悦禹 夏彦玲[1] LU Binshan;WANG Qianghui;ZHANG Qiang;GAO Yueyu;XIA Yanling(College of Wildlife Resources,Northeast Forestry University,Harbin 150040,China)

机构地区：[1]东北林业大学野生动物资源学院,哈尔滨150040

出　　处：《吉林农业大学学报》2019年第1期82-86,共5页Journal of Jilin Agricultural University

基　　金：中央高校基本科研业务费专项资金项目(2572014CA01);中国博士后科学基金项目(20110491124)

摘　　要：采用RT-PCR方法、分子克隆技术成功获得东北狍(Capreolus pygargus)PTN(pleiotrophin)基因cDNA序列。结果表明:获得的PTN基因cDNA序列长750 bp,共编码167个氨基酸,PTN蛋白分子质量为18.9 ku,理论等电点(pI)为9.66,是信号肽介导的一种亲水性蛋白质,其二级结构以无规则卷曲为主,其次是α-螺旋和β-折叠,分别占46.71%、32.34%和20.96%;生物信息分析发现,东北狍PTN基因在进化上较为保守,与其他物种同源性均>89%;构建的进化树表明,东北狍PTN基因与野牦牛、绵羊、野猪等偶蹄目动物亲缘关系较近,这一结果符合经典分类学规律;实时荧光定量RT-PCR分析显示,PTN基因在狍茸顶端组织不同生长层表达量存在显著差异,其中前软骨和间充质组织层的表达量明显高于皮肤和软骨组织层。In the present study, cDNA sequence of PTN gene from Northeastern roe deer was successfully cloned using RT-PCR technique and molecular cloning technology. Results showed that cDNA sequence of the PTN gene was 750 base pairs, which encoded 167 amino acids. The relative molecular weight of PTN protein was 18.9 ku, and theoretical isoelectric point(pI) was 9.66. PTN protein was a kind of hydrophilic proteins mediated by signal peptide and its secondary structure was given priority to with random curl, followed by alpha helix and beta-fold, accounting for 46.71%, 32.34% and 20.96%, respectively. Bioinformatics analysis found that the PTN protein of Northeastern roe deer was highly conserved in evolution, and homology with other species was over 89%. The evolutionary tree showed that the PTN protein of Northeastern roe deer had a close genetic relationship with artiodactyla animal, and the results conformed to the law of classical taxonomy. The results of Real-time RT-PCR showed that the expression level of PTN gene had significant difference among different tissues. The expression levels in mesenchymal and pre-cartilage were significantly higher than that in skin and cartilage.

关 键 词：东北狍 PTN基因 分子克隆 生物信息分析 实时荧光定量RT—PCR 

分 类 号：S825[农业科学—畜牧学] Q959.842[农业科学—畜牧兽医]