羊源多杀性巴氏杆菌ompW基因的克隆及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2019.01.004

Abstract

Abstract:

This study was aimed to clone ompW gene of Pasteurella multocida in goat and carry out bioinformatics analysis.According to the ompW gene sequence of Pasteurella multocida strain HN07 (accession No.:CP007040.1) in GenBank,one pair of primers were designed using DNAMAN 5.0 software,and the high-fidelity enzyme PrimeSTAR^® Max DNA Polymerase was selected 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 multocida in goat was highly homologous with porcine,bovine and avian-derived Pasteurella multocida,but not highly homologous to Pasteurella multocida in 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 27th 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.

Key words: Pasteurella multocida; ompW gene; clone; bioinformatics analysis

CLC Number:

S852.61⁺2

LI Baobao, ZHANG Zhenxing, HUANG Haifeng, ZHANG Mengmeng, ZHENG Yiying, WANG Chenqiang, ZHANG Luyin, AN Qi, YANG Xiaojian, NIE Xin, CAO Ruiyong, ZHU Shu, WANG Fengyang, DU Li. Cloning and Bioinformatics Analysis of ompW Gene from Pasteurella multocida in Goat[J]. , 2019, 46(1): 28-36.

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Cloning and Bioinformatics Analysis of ompW Gene from Pasteurella multocida in Goat

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