陆川猪脂酰辅酶A氧化酶1基因克隆及序列分析

摘要

The objective of this study was to clone the acyl-coenzyme A oxidase 1 (ACOX1) gene of Luchuan pig and analyze its genetic structure with bioinformatics.A pair of special primer was designed according to predicted sequence of pig ACOX1 gene in GenBank.The ACOX1 gene was amplified by RT-PCR, the physicochemical property and secondary structure of ACOX1 gene were systemically analyzed by bioinformatics techniques.The results showed that ACOX1 gene fragment included an 1 986 bp whole length CDS (coding 661 bp amino acids).The sequence multi-aligned results showed that Luchuan pig shared 99.5%, 85.5%,87.1%,66.3%,75.6%,84.0%,82.3%,81.1% and 69.1% of similar nucleotide sequence with that of pig,human, zebrafish, chicken, rhesus monkeys, mice, rat and frog, respectively.The prediction of ACOX1 secondary structure showed that Luchuan pig ACOX1 had no signal peptide and transmembrane structure.This study suggested that the whole CDS sequence of ACOX1 gene was successfully cloned in Luchuan pig, and the cloning and analysis of ACOX1 gene provided an important foundation for further studying on the fatty deposition and lipornetabolism of ACOX1 gene in Luchuan pig.%本研究旨在对陆川猪脂酰辅酶A氧化酶1(acyl-coenzyme A oxidase 1,ACOX1)基因进行克隆及生物信息学分析.根据GenBank中猪ACOX1基因序列设计特异性引物,利用RT-PCR技术对陆川猪ACOX1基因进行克隆、测序和生物信息学分析.结果表明,陆川猪ACOX1基因CDS全长1 986 bp,编码661个氨基酸,陆川猪ACOX1基因与猪、牛、人、斑马鱼、鸡、猕猴、大鼠、小鼠、爪蟾序列同源性分别为99.5%、85.5%、87.1%、66.3%、75.6%、84.0%、82.3%、81.1%和69.1%.系统进化树分析结果表明,ACOX1基因在不同物种及进化的过程中具有高度保守性,蛋白质二级结构预测结果表明,陆川猪ACOX1蛋白没有信号肽,没有形成跨膜结构.本研究成功克隆了陆川猪ACOX1基因,为阐明其在陆川猪脂肪沉积及代谢方面的调控研究奠定了理论基础.