中国畜牧兽医2019，Vol.46Issue(1)：28-36,9.DOI:10.16431/j.cnki.1671-7236.2019.01.004
羊源多杀性巴氏杆菌ompW基因的克隆及生物信息学分析
Cloning and Bioinformatics Analysis of ompW Gene from Pasteurella multocidain Goat
摘要
Abstract
This study was aimed to clone ompW gene of Pasteurella multocidain goat and carry out bioinformatics analysis.According to the ompWgene sequence of Pasteurella multocidastrain HN07 (accession No.:CP007040.1) in GenBank, one pair of primers were designed using DNAMAN 5.0software, and the high-fidelity enzyme PrimeSTAR? Max DNA Polymerase was se-lected for PCR reaction to obtained the target gene fragment, the nucleotide and amino acid sequences of ompW gene were analyzed by bioinformatics softwares.The results showed that the length of PCR product was about 615 bp, which encoded 204 amino acids.The nucleotide homology analysis result showed that ompW gene of Pasteurella multocidain goat was highly homologous with porcine, bovine and avian-derived Pasteurella multocida, but not highly homologous to Pasteurella multocidain rabbit.The phylogenetic tree result showed that the relationship was the closest between ompW gene of Pasteurella multocida in goat and porcine.The bioinformatics analysis results showed that the molecular formula of ompW protein was C1007H1567N257O283S3, the molecular weight was 21.90 ku, the theoretical isoelectric point (pI) was 9.16, which was a basic protein with a hydrophobic index of 96.57 and a total average hydrophobicity (GRAVY) of 0.173 (>0) , it belonged to a hydrophobic protein.The first 21 amino acids were signal peptides, the 5th to 27 th amino acid regions had a transmembrane region, there were N-glycosylation and phosphorylation sites, no O-glycosylation site, and there were multiple B cell, CTL cell and Th cell epitopes.The alpha helix, extended chain, beta turn and random coil of the secondary structure accounts for 17.65％, 35.29％, 3.92％ and 43.14％, respectively.The tertiary structure was a beta barrel monomer belonging to one of the members of the outer membrane protein family.These results provided a theoretical basis for further elucidating the mechanism of anti-host immune stress and the development of vaccines in the process of infecting host of Pasteurella multocida.关键词
多杀性巴氏杆菌/ompW基因/克隆/生物信息学分析Key words
Pasteurella multocida/ompW gene/clone/bioinformatics analysis分类
农业科技引用本文复制引用
李宝宝,聂鑫,曹瑞勇,朱姝,王凤阳,杜丽,张振兴,黄海峰,张萌萌,郑义盈,王成强,章泸尹,安琪,杨小健..羊源多杀性巴氏杆菌ompW基因的克隆及生物信息学分析[J].中国畜牧兽医,2019,46(1):28-36,9.基金项目
海南省重大科技计划项目(ZDKJ2016017-01) （ZDKJ2016017-01）
国家肉羊产业技术体系(CARS-38) （CARS-38）
中央引导地方科技发展专项资金项目(ZY2017HN07) （ZY2017HN07）
