奶山羊USF1基因的克隆与组织表达分析

Cloning, Sequence Analysis and Tissue Expression of USF1 Gene in Dairy Goat

为了获得奶山羊(Capra hircus)USF1基因序列并分析其在不同泌乳时期乳腺组织中的表达规律,从奶山羊乳腺组织中克隆得到USF1基因序列(Gen Bank登录号:MF953285),CDS区序列933 bp,编码310个氨基酸。通过序列比对发现,山羊与牛(Bos taurus)、猪(Sus scrofa)和人(Homo sapiens)的USF1基因核苷酸和氨基酸序列相似性均较高,为94%以上。蛋白质结构预测发现,USF1蛋白质结构为典型的螺旋-环-螺旋结构;其蛋白不含跨膜结构域,亚细胞定位在细胞核内。实时荧光定量PCR分析表明,USF1基因在奶山羊泌乳前期、盛期和中期的乳腺组织中m RNA表达量均显著高于干奶期(P<0.05)。因此,USF1可能参与奶山羊乳腺的泌乳过程。该研究可为进一步揭示USF1基因在奶山羊乳腺组织中的功能及对脂质代谢的调控作用提供基础资料。

The aim of this study was to clone USF1 and analyze its m RNA expression level in mammary gland of dairy goat(Capra hircus)in different lactation periods.Coding sequence(CDS)of USF1 was amplified(Gen Bank accession number:MF953285)from mammary gland of dairy goat. The results showed that the CDS length of USF1 gene was 933 bp,coding 310 amino acids. The homology analysis of dairy goat USF1 gene indicated that the nucleotides and amino acids had high similarity with bovine(Bos taurus),porcine(Sus scrofa),and human(Homo sapiens),and it reached up to more than 94%. The protein structure analysis showed that USF1 was the basic helix-loop-helix structure. There was notransmembrane structure in USF1 protein. Subcellular localization analysis showed USF1 located in the nucleus.The real-time fluorescence quantitative PCR analysis showed that the m RNA expression levels of USF1 in mammary gland of dairy goat in the lactation were significantly higher than that in dry period(P<0.05). Taken together,these results indicate that USF1 may participate in the lactation process of mammary gland in dairy goat,and provide some basic information for further revealing of the USF1 function and the regulation of lipid metabolism by USF1 in mammary gland.