马鹿CDK8基因生物信息学分析与组织表达

Bioinformatics Analysis and Tissue Expression of CDK8 Gene in Red Deer

为探究细胞周期蛋白依赖性蛋白激酶8基因(CDK8)在马鹿(Cervus elaphus)鹿茸的皮、间充质、前软骨和软骨组织中的m RNA表达水平和该基因编码蛋白的生物学特性,采用PCR方法克隆马鹿CDK8基因编码区(CDS)全长序列,并连入p MD-18T载体,测得核苷酸序列,进行相似性比对和系统进化树构建;利用生物信息学方法分析CDK8蛋白的组成、结构和理化性质;利用实时荧光定量PCR技术检测CDK8基因m RNA在鹿茸组织中的表达水平。结果表明:马鹿CDK8的CDS全长1254 bp,共编码417个氨基酸;所编码的蛋白相对分子质量为47744.11,理论等电点(p I)为7.28,是一种无信号肽、定位于细胞核的亲水性不稳定蛋白,共38个磷酸化位点,蛋白的二级结构以无规则蜷曲和α螺旋为主。CDK8的mRNA在茸皮组织中表达量最高,其后依次是间充质、软骨和前软骨。CDK8是转录机器中介体复合物的重要组成部分,研究克隆获得马鹿CDK8的CDS全长序列,可为了解鹿茸再生过程中CDK8的作用机制与功能提供重要的基础数据。

In order to explore the mRNA expression level of cyclin-dependent protein ki⁃nase 8 gene(CDK8)in various tissues of red deer(Cervus elaphus)antler velvet and the biological characteristics of the protein encoded by the gene,the full-length coding sequence(CDS)of cyclin-dependent pro⁃tein kinase 8 gene of red deer was cloned by PCR and connected to pMD-18T vector to measure nucleotide sequences.Thereafter,similarity alignment and phylogenetic tree construction could be carried out,and the resulting nucleotide se⁃quences was further used for similarity comparison and phylogenetic tree construction.The composition,structure and physicochemical properties of CDK8 protein were analyzed by bioinformatics methods.Real-time PCR was used to detect the expression level of CDK8 gene mRNA in antler tissues.The results showed that the coding region of red deer CDK8 was 1,254 bp in length,and a total of 417 amino acids were encoded.The encoded protein has a relative molecular mass of 47,744.11 and a theoretical isoelectric point pI of 7.28,which is a hydrophilic unstable protein with no signal peptide and localized to the nucleus with a total of 38 phosphorylation sites,and the secondary structure of the protein is dominated by irregular curls andαhelixes.The expression level of CDK8 mRNA is the highest in furry tissue,followed by mesenchymal,then cartilage,and the lowest is in precartilage.CDK8 is an important part of the transcription machine the mediator tran⁃scriptional regulatory complex.In this study,the full-length coding sequence of red deer CDK8 was cloned for the first time,which can provide important reference data for us to understand the mechanism and function of CDK8 during antler regen⁃eration.