广西巴马小型猪MCP-1基因克隆及序列分析

Cloning and analysis of MCP-1 gene in Guangxi Bama mini-pig

【目的】研究广西巴马小型猪单核细胞趋化蛋白-1(MCP-1)基因CDs区的序列特点,为建立广西巴马小型猪动脉粥样硬化模型及揭示动脉粥样硬化的发生、发展机理提供理论依据。【方法】从广西巴马小型猪动脉血管组织中扩增MCP-1基因CDs区全长序列,与人类及其他易建立动脉粥样硬化模型动物的序列进行比对分析,并预测其蛋白质信号肽位点及结构域。【结果】广西巴马小型猪MCP-1基因CDs区全长300 bp,编码99个氨基酸;与人类MCP-1基因CDs区(S71513.1)的同源性最高,为84.4%。广西巴马小型猪MCP-1蛋白的前23位氨基酸为信号肽,后76位氨基酸为成熟肽,等电点(pI)9.51,蛋白质分子量10976.04kDa,信号肽位置有一段跨膜结构;广西巴马小型猪与人类MCP-1蛋白在形成二级结构时,保守半胱氨酸参与形成二硫键的位置一致,分别在第11、12、36和52号(除信号肽片段)位上排列为Cys-Cys,且是第11号位与第36号位形成二硫键,第12号位与第52号位形成二硫键。【结论】以广西巴马小型猪构建动脉粥样硬化模型时,MCP-1基因表达量可作为判断动脉血管疾病的一个辅助指标。

[Objective]The sequence features of monocyte chemotactic protein 1 （MCP-1） gene of Guangxi Bama mini-pig were researched to provide theoretical references for animal models construction and study on the pathogene- sis of atherosclerosis. [Method]The complete coding sequence （CDs） of MCP-1 gene was cloned from Guangxi Ba- ma mini-pigs＇ vascular tissue. This sequence was compared with that of human and that of other animals which were used to construct atherosclerosis models. The Bama mini-pig MCP-1 protein signal peptide and domains were predicted. [Result]The study results showed that Guangxi Bama mini-pig MCP-1 gene CDs had 300 bp in length, which en- coded 99 amino acids. Guangxi Bama mini-pig MCP-1 gene CDs had the greatest homology with that of human （S71513.1）, which was 84.4%. The start 23 amino acids of the MCP-1 protein were the signal peptide, and the lat- er 76 amino acids were mature peptide. This protein isoelectrie point （pI） was 9.51, and its protein molecular weight was 10976.04 kDa, and there was a transmembrane in the signal peptide, which was the same as human MCP-I protein secondary structure. Guangxi Bama mini-pig MCP-1 protein contained 2 disulfide bonds, which were the 11th （except of the signal peptide） Cys linking with 36th Cys, and 12th Cys linking with 52th Cys. [Conclusion] The MCP- 1 gene expression could be used as an auxiliary index for judging occurrence of arterial vascular disease, when con- structing the Guangxi Bama mini-pig atherosclercosis models.