摘要
为了为鹿茸生长发育机制及CDKN2C基因在鹿茸生长发育中所发挥的重要功能研究提供理论依据和技术支持,试验根据GenBank中公布的牛、绵羊的CDKN2C基因序列设计引物,采用PCR扩增和分子克隆技术,首次从梅花鹿鹿茸顶端组织中克隆出包含CDKN2C全部编码区的基因序列,利用生物信息学方法对该基因核苷酸序列以及编码氨基酸序列的结构特征进行初步预测分析,并基于该基因氨基酸序列构建了系统进化树。结果表明:梅花鹿CDKN2C基因CDS区全长507 bp,编码168个氨基酸,系统进化树结果显示其与大羚羊、野骆驼、牛以及马的亲缘关系较近,其编码蛋白质的相对分子质量为18 325.67,一级结构中亮氨酸和丙氨酸含量较多,分别占比14.3%和13.1%,为亲水非分泌性蛋白,无信号肽。结构域预测发现第46~127位氨基酸残基之间存在Ank;结构域。二级结构分析显示该蛋白α-螺旋占47.62%,延伸链占12.50%,自由卷曲占39.88%。三级结构预测显示其结构与牛更为相似。该蛋白含有2个丝氨酸和3个苏氨酸磷酸化位点,分别位于氨基酸序列的第23,154位和第88,126,139位,没有发现N-糖基化位点以及O-糖基化位点。
The primers were designed according to the CDKN2 C gene sequence of cattle and sheep published in GenBank.The c DNA sequence containing all coding regions of CDKN2 C was cloned from the top tissue of deer antler for the first time by PCR amplification and molecular cloning technology.The structural characteristics of the nucleotide sequence and amino acid sequence coded were preliminarily predicted and analyzed by bioinformatics,and the phylogenetic tree was constructed based on the amino acid sequence of the gene.The results showed that the full length of CDKN2 C gene coding sequence of sika deer was 507 bp,encoding 168 amino acids.The phylogenetic tree showed that the CDKN2 C gene was closely related to the Oryx dammah,Camelus ferus,Bos taurus and Equus caballus.The relative molecular weight of the protein was 18325.67.The content of leucine and alanine in primary structure is the highest,accounting for 14.3%and 13.1%respectively,which is a hydrophilic non-secretory protein without signal peptide.The prediction of the domain shows that there was a Ank_2 domain between the amino acid residues at positions 46 to127.The secondary structure showed that the protein was composed of random coil,alpha-helix,extended chain and free curl,which account for 47.62%,12.50%,39.88%respectively.The tertiary structure prediction showed that the structure was more similar to that of Bos taurus.The protein contains 2 serine and 3 threonine phosphorylation sites,located 23,154 and 88,126,139 of the amino acid sequence respectively,no N-glycosylation sites and O-glycosylation sites were found.
作者
李金跃
刘嫣然
赵晨曦
于梦佳
刘泳彤
黄雨馨
夏彦玲
LI Jinyue;LIU Yanran;ZHAO Chenxi;YU Mengjia;lIU Yongtong;HUANG Yuxin;XIA Yanling(School of Wildlife and Natural Reserve,Northeast Forestry University,Harbin 150040,China)
出处
《黑龙江动物繁殖》
2021年第5期1-7,共7页
Heilongjiang journal of animal reproduction
基金
黑龙江省大学生创新项目(201910225307)
