陆川猪脂联素基因的克隆及生物信息学分析

The Cloning and Bioinformatics Analysis of AdipoQ Gene in Luchuan Pig

脂联素(AdipoQ)是脂肪组织分泌的一种有活性的物质,其在调节脂肪代谢、脂肪沉积以及对抗胰岛素抵抗、炎症反应、动脉粥样硬化等方面发挥着重要的作用。本研究以Gen Bank家猪脂联素基因序列为参考,利用Oligo7自行设计一对引物,以广西陆川猪皮下脂肪组织cDNA为模板,克隆获得了AdipoQ基因编码区序列(CDS)。序列分析结果显示,陆川猪AdipoQ基因CDS区长为732 bp,可编码242个氨基酸,分子量为26 400.7 Da。信号肽预测显示陆川猪Adipo Q蛋白含有编码17个氨基酸的信号肽,这一预测结果也印证了前人的研究,进一步证实了脂联素为一种分泌蛋白的说法。与GenBank已公布的长白猪(DQ164208.1)、大白猪(DQ164207.1)、广西巴马小型猪(JF730719.1)、梅山猪(DQ164209.1)这四种猪AdipoQ基因CDS区进行序列比较,发现不同猪种间略有差异,而同源性均达98.9%以上。本研究还对克隆获得的陆川猪AdipoQ基因编码蛋白进行了高级结构预测,为后期研究AdipoQ基因调控陆川猪脂肪沉积和肌肉生长发育的分子机制提供理论基础。

Adiponectin protein is a kind of active substance secreted by adipose tissue, which play an important role in the aspect of regulating metabolism, insulin resistance, anti-inflammatory, and resistance to atherosclerosis and fat deposition. In this study, we designed a pair of AdipoQ primer by Oligo 7, according to the pig A dipoO gene sequence in GenBank. And then, we obtained the coding sequence （CDS） of AdipoQ by cloning with the cDNA of subcutaneous adipose tissue in Luchuan pig. After sequence analyzing, we concluded that the CDS length of AdipoQ for Luchuan pig was 732 bp, encoding 242 amino acids, and its molecular weight was 26 400.7 Da. The signal peptide prediction of Luchuan pig AdipoQ showed it contained signal peptide and encoded 17 amino acids, which confirmed the previous research result that AdipoQ was a secreted protein. Among Luchuan pig, Landrace pig （DQ 164208.1）, Large White pig （DQ 164207.1）, Guangxi B ama mini-pig （JF730719.1 ） and Meishan pig （DQ 164209.1）, the sequence homology of AdipoQ gene were more than 98.9%, and there was slight difference in different breeds. The study also made a prediction about the advanced structure of Luchuan pig＇s A dipoO gene, which laid the foundation for further molecular mechanism research between AdipoQ expression and fat deposition, muscle development.