奶山羊CIDEa基因的克隆、序列分析与组织表达

为获得奶山羊CIDEa基因序列并检测其在乳腺组织中的表达,采用RT-PCR技术从奶山羊乳腺组织扩增CIDEa基因序列,利用生物信息学软件对其进行预测分析,并采用荧光定量PCR检测CIDEa在干奶期和不同泌乳时期乳腺组织中的表达。结果表明:通过克隆测序得到奶山羊CIDEa基因CDS区660 bp,编码219个氨基酸,与GenBank公布的绵羊、牛、人和猪的核苷酸序列同源性分别为99%、96%、85%、85%。奶山羊CIDEa蛋白属于碱性、不稳定亲水蛋白,不存在跨膜结构和信号肽;蛋白主要定位在细胞质、线粒体和细胞核。CIDEa蛋白结构主要由α螺旋、β折叠和不规则卷曲组成。奶山羊CIDEa在泌乳前期、盛期和中期乳腺组织中表达量均显著高于干奶期(P<0.05)。说明CIDEa可能参与奶山羊乳腺的泌乳过程,为进一步研究其在奶山羊乳腺组织中的功能及对乳品质的调控作用奠定基础。

Cloning,sequence analysis and tissue expression of the CIDEa gene in dairy goat

In order to obtain the cell death-inducing DNA fragmentation factor 45-like effector A(CIDEa)gene sequence of dairy goat and to detect its expression in mammary gland ofof the animal,the sequence of the CIDEa gene was amplified from the mammary gland of dairy goat by RT-PCR,and was analyzed using a bioinformatics software.The expression of CIDEa in different lactation periods was detected by fluorescence quantitative PCR.The results showed that the complete CDS of the CIDEa gene was 660 bp,encoding 219 amino acids,which were in 99%,96%,85%,85%homologous with that of Ovis aries,Bos taurus,Homo sapiens,and Sus scrofa.The CIDEa protein belonged with the alkaline unstable hydrophilic protein,and it had no transmembrane structure or signal peptide.The protein was mainly located in the cytoplasm,mitochondria and nuclear.Its structure consisted ofα-helices,β-foldings,and random coils.The expression of the CIDEa gene in the lactation period was significantly higher than that in the dry period(P<0.05).These results indicated that CIDEa might be involved in the lactation process of dairy goat,which laid a foundation for further research on its function in the mammary gland and its regulation effect on the milk quality,of dairy goat.