白洗猪PID1基因克隆及序列分析

本研究旨在对白洗猪磷酸酪氨酸互作结构域1 (phosphotyrosine interaction domain containing 1,PID1)基因进行克隆和生物信息学分析。采用Nest PCR及T克隆技术对白洗猪PID1基因进行克隆测序,运用生物学分析软件分析其结构功能及其在种内及种间的遗传进化关系。结果表明,白洗猪PID1基因CDS区全长654 bp,编码217个氨基酸,白洗猪与山东莱芜猪、广西陆川猪PID1蛋白氨基酸同源性为98.2%与97.7%;进化树分析结果表明白洗猪与两个猪种间遗传关系相对较远,种间比对黄牛、牦牛、猕猴、小鼠、眼镜王蛇、人、原鸡、非洲爪蟾、大鼠和斑马鱼PID1蛋白氨基酸同源性依次为96.6%、96.6%、96.3%、95.0%、93.9%、91.7%、90.8%、88.2%、69.7%和67.3%;系统进化树分析表明PID1基因在多物种之间的进化高度保守,结构功能分析表明白洗猪PID1基因功能区主要是编码链氨基酸C端的PTB结构域。本研究成功克隆了白洗猪PID1基因,为探究其对白洗猪肌内脂肪沉积方面的影响及为白洗猪种资源开发利用奠定理论基础。     

Cloning and Sequence Analysis of PID1 Gene in Baixi Pig

In order to study phosphotyrosine interaction domain containing (PID1) gene of Baixi pig, it was amplified and sequenced by Nest PCR and T clone technology.Then the functions and the genetic evolutionary relationships of the gene and its predicted protein were analyzed by bioinformatics software.The results showed that the whole CDS region of PID1 gene was 654 bp, which encoded 217 amino acids.The results of sequence alignments showed that Baixi pig shared 98.2% and 97.7% similarities of amino acid with which of Shandong Laiwu pig and Guangxi Luchuan pig.The phylogenetic tree indicated that Baixi pig kept away from these two pigs.The results of sequence alignments among species showed that Baixi pig shared 96.6%, 96.6%, 96.3%, 95.0%, 93.9%, 91.7%, 90.8%, 88.2%, 69.7% and 67.3% similarities of amino acid with which of Bos taurus, Bos grunniens, Macaca mulatta, Mus musculus, Ophiophagus hannah, Homo sapiens, Gallus gallus, Xenopus laevis, Rattus norvegicus and Danio rerio.Phylogenetic analysis indicated that PID1 gene was highly conserved in the process of evolution of different species.The protein structure analysis results showed that the mainly function region of PID1 gene was PTB structural domain, which was located in the C-terminal sequence of the PID1 protein.In this study, we successfully cloned PID1 gene of Baixi pig, which laid the foundation for further study in intramuscular fat deposition and development of resources.