梅花鹿PTN基因cDNA克隆及表达分析  

cDNA cloning and expression analysis of PTN gene from sika deer

作　　者：崔薇 陈楠 苗震 李和平[1] 刘伟石[1] 夏彦玲[1] CUI Wei;CHEN Nan;MIAO Zhen;LI Heping;LIU Weishi;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：《黑龙江畜牧兽医》2019年第7期129-132,共4页Heilongjiang Animal Science And veterinary Medicine

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);东北林业大学大学生创新训练项目(201710225216)

摘　　要：为了探究多效生长因子(PTN)基因在鹿茸生长发育中的调控作用,试验采用分子克隆技术得到包括PTN基因全部编码区的cDNA序列,并结合生物信息学方法、实时荧光定量PCR技术对其进行分析。结果表明:获得的PTN基因cDNA序列长度为750 bp,共编码168个氨基酸;PTN蛋白分子质量为18.93 ku,理论等电点为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47.62%、32.74%和19.64%;梅花鹿PTN基因在进化上较为保守,与白尾鹿、野猪、牛等偶蹄目动物亲缘关系较近,与斑纹海豚、豚鼠、人亲缘关系较远;实时荧光定量PCR分析显示,PTN基因在不同生长时期的鹿茸顶端间充质组织中表达差异显著,表达量在生长中期明显高于前期和后期。In order to study the regulating effect of PTN gene on the growth and development of sika deer antler,the cDNA sequence including complete coding region of PTN gene was obtained by molecular cloning technology and analyzed by RT-qPCR and bioinformatics methods. The results showed that cDNA sequence of the PTN gene was 750 bp,which encoded 168 amino acids. The relative molecular weight of PTN protein was 18. 93 ku,and theoretical isoelectrisc point ( PI) was 9. 66. PTN protein was a kind of hydrophilic proteins mediated by signal peptide and its secondary structure was made up of random curl,followed by alpha helix and beta-fold,accounted for 47. 62%,32. 74% and 19. 64% re-spectively. Bioinformatics analysis found that the PTN protein of sika deer was highly conserved in evolution. The evolutionary tree showed that the PTN protein of sika deer had a close genetic relationship with Odocoileus virginianus texanus,Sus scrofa and Bos taras. It was far genetic re-lationship with Lagenorhynchus obliquiden,Cavia porcellus and Homo sapiens. The result of RT-qPCR showed that there was a significant differ-ence in the expression level of PTN gene in the antler mesenchymal tissue at different growth stages. The PTN gene expression levels in the me-dium term were significantly higher than in the early and late stages.

关 键 词：梅花鹿 PTN 基因 鹿茸 间充质组织 实时荧光定量 PCR 生物信息学分析 

分 类 号：S825[农业科学—畜牧学]