摘要： 旨在克隆牦牛诱导细胞凋亡DNA片段化45样效应因子A（CIDEA）基因序列、分析其生物学特性及检测CIDEA基因在牦牛不同组织及脂肪细胞分化中的表达模式。以金川牦牛皮下脂肪组织的cDNA为模板，采用PCR技术克隆CIDEA基因的CDS序列（Coding sequence），对CDS区进行相关的生物信息学分析；设计特异引物，检测该基因在金川牦牛肺脏、心脏、肾脏、脂肪等组织的表达情况；同时分离原代前体脂肪细胞，分析CIDEA基因在脂肪细胞分化过程中的表达趋势。结果表明，金川牦牛CIDEA基因的序列长度为696 bp，其中CDS序列为684 bp，编码227个氨基酸；金川牦牛CIDEA基因与普通牛的同源性最高，核苷酸序列同源性为94.14%，氨基酸序列同源性为99.54%，与鸡的同源性最低，核苷酸序列同源性仅为63.38%，氨基酸序列同源性仅为59.05%，利用MEGA 5.0软件构建进化树显示金川牦牛与牛的亲缘关系最近，与鸡的亲缘关系最远。蛋白理化性质分析结果表明，CIDEA蛋白是一个不稳定的碱性亲水蛋白，不存在跨膜结构与信号肽；二级结构预测显示CIDEA蛋白中α螺旋占33.92%，无规则卷曲占51.98%。实时荧光定量PCR结果表明，CIDEA基因在金川牦牛脂肪组织中表达量最高，肺脏中表达量最低；在金川牦牛脂肪细胞分化过程中呈现上升的趋势。由此可推测金川牦牛CIDEA基因可能参与牦牛脂肪细胞分化与脂肪沉积。

关键词: 金川牦牛, CIDEA基因, 基因克隆, 时序表达, 脂肪组织, 脂肪细胞

Abstract: The aim of this study was to clone the gene sequence of cell death-inducing DNA fragmentation factor 45-like effector A (CIDEA) in yak, analyze its biological characteristics, and detect the expression pattern of CIDEA in yak different tissues and during the differentiation of adipocytes. The CDS sequence of CIDEA was cloned by PCR, using Jinchuan yak subcutaneous adipose tissue cDNA as a template, and the biological characteristic was analyzed;specific primers were designed to detect the expression of CIDEA in various Jinchuan yak tissues, including lung, heart, kidney, fat and other tissue of Jinchuan yak. And primary adipocytes were isolated to analyze the expression trend of CIDEA during adipocyte differentiation. The results showed that the length of Jinchuan yak CIDEA was 696 bp, of which CDS was 684 bp, encoding 227 amino acids. CIDEA of Jinchuan yak had the highest homology to that of Bos taurus, representing that the nucleotide identity was 94.14% and the amino acid homology was 99.54%, and the lowest homology to that of Gallus gallus, representing that the nucleotide identity only was 63.38% and the amino acid homology only was 59.05%. Also, phylogenetic tree constructed by MEGA 5.0 software showed that similarity was highest between Jinchuan yak and cattle, and similarity was lowest between Jinchuan yak and chicken. Moreover, the physical and chemical properties of protein were predicted that CIDEA protein was an unstable basic hydrophilic protein without transmembrane structure and signal peptide. The secondary structures of CIDEA protein mainly consisted of 33.92% α-helix and 51.98% random coils. Real-time quantitative PCR results showed that the expression quantity of CIDEA was highest in Jinchuan yak adipose tissue and was lowest in lung. And the differentiation of adipocytes in Jinchuan yak showed an increasing trend. It is concluded that the expression of CIDEA might involve in adipose differentiation and adipose deposition in yaks.

Key words: Jinchuan yak, CIDEA, Gene cloning, Temporal-expression, Adipose tissue, Adipocyte

中图分类号: 

• Q78

• S823