梅花鹿S100A16基因的克隆及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2020.01.004

摘要/Abstract

摘要： 为探索梅花鹿（Cervus nippon）S100A16基因序列及生物学特性，本研究根据GenBank数据库中牛、绵羊S100A16基因序列设计引物，以梅花鹿鹿茸顶端组织cDNA为模板，采用RT-PCR技术和分子克隆技术成功获得梅花鹿S100A16基因的cDNA序列。生物信息学分析发现，梅花鹿S100A16基因CDS区全长312 bp，编码103个氨基酸；蛋白含有11个磷酸化位点，有跨膜结构域，无信号肽，为在细胞内发挥作用的稳定蛋白；蛋白仅在C端含有S100蛋白家族经典的EF螺旋结构域，由12个氨基酸组成，N端EF螺旋结构域由15个氨基酸组成；蛋白C端含有FGF-1蛋白结合位点；梅花鹿S100A16蛋白的二级结构主要由α-螺旋和无规则卷曲构成；三级结构显示该蛋白有2个Ca²⁺结合位点；梅花鹿S100A16蛋白氨基酸序列与东欧马鹿同源性最高，为100%，与其他部分物种S100A16蛋白氨基酸序列构建系统进化树，分析表明S100A16基因在进化上比较保守，符合功能基因的特点。研究结果为进一步揭示梅花鹿S100A16基因的功能及表达机制提供依据。

关键词: 梅花鹿; S100A16基因; 分子克隆; 生物信息学分析

Abstract: In order to explore the S100A16 gene sequence and biological characteristics of Sika deer (Cervus nippon),primers were designed according to the S100A16 gene sequence of cattle and sheep in GenBank database,and the cDNA of S100A16 gene in Sika deer was successfully obtained by RT-PCR and molecular cloning technology with the top tissue cDNA of sika deer antler as template.Bioinformatics analysis showed that the total length of CDS region of S100A16 gene in Sika deer was 312 bp,encoding 103 amino acids;the protein contained 11 phosphorylation sites,transmembrane domain,and no signal peptide,which was the stable protein that played a role in the cell;The protein only contained the EF-hand of S100 protein family in the C-terminal,which was composed of 12 amino acids,and the EF-hand at the N-terminal was composed of 15 amino acids.The second-order structure of S100A16 protein in Sika deer was mainly composed of alpha-helix and ranchorn coil;the third-order structure showed that the protein had two Ca²⁺ binding sites;the amino acid sequence of S100A16 protein in Sika deer had the highest homology with Eastern European red deer,which was 100%,and the phylogenetic tree was constructed with the amino acid sequence of S100A16 protein in some other species.S100A16 gene was relatively conservative in evolution and conformed to the characteristics of functional genes.The results provided the basis for further revealing the function and expression mechanism of S100A16 gene in Sika deer.

Key words: Sika deer; S100A16 gene; molecular cloning; bioinformatics analysis

中图分类号:

Q785

陈继乾, 夏彦玲, 高悦禹, 刘伟石. 梅花鹿S100A16基因的克隆及生物信息学分析[J]. 中国畜牧兽医, 2020, 47(1): 28-36.

CHEN Jiqian, XIA Yanling, GAO Yueyu, LIU Weishi. Cloning and Bioinformatics Analysis of S100A16 Gene in Sika Deer[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(1): 28-36.

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