东北马鹿ELK1基因CDS序列克隆及其生物信息学分析

Cloning and bioinformatics analysis of the CDS sequence of ELK1 gene in Northeastern Red deer

为探究东北马鹿ELK1基因CDS序列信息,试验进行了mRNA反转录、CDS序列扩增,测定ELK1基因CDS序列,克隆并构建蛋白表达载体,利用生物信息学方法对该基因编码的蛋白质结构及理化性质等进行了预测与分析。结果表明:东北马鹿ELK1基因转录产物CDS序列全长为1323 bp(GenBank登录号为ON212657),编码440个氨基酸,分子量为46200.42 u。核酸序列具有进化保守性,与牛ELK1基因同源性最高,为97.99%。ELK1多肽链具有亲水性,没有分泌信号肽结构,等电点为5.81,蛋白质为不稳定蛋白。ELK1蛋白的二级结构由无规则卷曲、α-螺旋、延伸链、β-折叠组成,存在ETS DNA结合域,亚细胞定位于细胞核。

In order to investigate the sequence information of CDS sequence of ELK1 gene, in the experiment, mRNA reverse transcription and CDS region amplification were used to determine the sequence of CDS sequence of ELK1 gene, which was cloned and constructed into the eukaryotic expression vector. Bioinformatics methods were used to predict and analyze the structure and physicochemical properties of the protein encoded by the gene. The results showed that the CDS sequence of ELK1 gene was 1 323 bp in length(GenBank accession number ON212657) and encoded 440 amino acids with a molecular mass of 46 200.42 u. The nucleic acid sequence was evolutionarily conserved and showed the highest homology with bovine ELK1 gene(97.99%). The ELK1 polypeptide chain was hydrophilic, without secretory signal peptide structure, and had an isoelectric point of 5.81;the protein was unstable. The secondary structure of ELK1 protein consisted of irregularly coiled, α-helix, extended chain, β-fold, having ETS structural domain, and was located in the nucleus under subcellular localization.