摘要摘 要 YWHAEA (assignment of the human 14-3-3 epsilon isoform)基因编码14-3-3ε蛋白，此蛋白在发育的生茸区骨膜中特异性高表达。为了克隆梅花鹿(Cervus nippon) YWHAE基因编码区序列，预测其蛋白结构和功能，并研究其在梅花鹿发育过程中的表达特性。本研究以梅花鹿生茸区骨膜为材料，采用RT-PCR技术克隆梅花鹿YWHAE基因编码区序列，利用生物信息学手段预测其结构与功能。实验结果表明，梅花鹿YWHAE基因编码区全长768 bp (GenBank No. KY797653)，编码255个氨基酸，氨基酸水平上与牛(Bos taurus)相似性高达99.6%；氨基酸序列分析表明，鹿YWHAE基因编码的14-3-3蛋白相对分子量为29 163.86 D，主要定位在细胞质内，属于水溶性蛋白；预测鹿14-3-3蛋白具有19个磷酸化位点、不存在糖基化位点；该蛋白的二级结构主要为螺旋结构，其三级结构呈现出递进螺旋状态。预测14-3-3蛋白的结构特点，本研究为探究14-3-3蛋白与其他蛋白的相互作用提供理论依据，同时为进一步研究鹿茸发育的调控机理提供基础依据。
Abstract：Abstract Assignment of the human 14-3-3 epsilon isoform (YWHAE) gene encodes 14-3-3ε protein, which highly specifically expresses during antler development of sika deer(Cervus nippon). The study was to clone the YWHAE gene's cDNA coding domain sequence of sika deer and predict its structure, function and expression profiles. Using antlerogenic periosteum as materials, the sika deer's YWHAE CDS was cloned by RT-PCR and its protein structure and functions were predicted through several bioinformational tools as well. The results showed that the sika deer's YWHAE CDS consisted of 768 base pair (GenBank No. KY797653) that encoded 255 amino acids residues, and the amino acid homology of deer YWHAE shared 99.6% identity with bovine (Bos taurus). The analysis of amino acid sequence revealed that the sika deer YWHAE encoded 14-3-3 protein and its relative molecular weight was 29 163.86 D. Subcellular location of 14-3-3 protein was primarily in the cytoplasm, and it belonged to the water-soluble protein. It was predicted that the 14-3-3 protein contained 19 phosphorylation sites, and there was no glycosylation sites. The secondary structure of the protein mainly had helixs, while the tertiary structure of domain area showed a helical state. The study predicts the characteristics of the 14-3-3 protein structure and provides basis theory on exploring the 14-3-3 proteins interacting with other proteins. Furthermore, it provides a basis for further studying on the regulation mechanism of antler development.
