猪多杀性巴氏杆菌OmpW、TbpA蛋白的原核表达及免疫原性研究

doi:10.16431/j.cnki.1671-7236.2023.04.021

Abstract

Abstract: 【Objective】 This study was aimed to investigate the immunogenicity of OmpW and TbpA proteins of swine Pasteurella multocida and explore their potential as vaccine candidate antigens.【Method】 The similarity of amino acid sequences of OmpW and TbpA proteins in different serotypes of swine Pasteurella multocida were compared by NCBI.Further bioinformatics analysis of OmpW and TbpA proteins were carried out.Based on this,the recombinant expression strains pET28a-OmpW-BL21 and pET28a-TbpA-BL21 were successfully constructed,which were purified and mixed with aluminum hydroxide adjuvant to prepare subunit vaccine.After immunizing mice,the immunogenicity of OmpW and TbpA proteins was evaluated using swine Pasteurella multocida GX-PmD4 strain to challenge.【Result】 The results of similarity analysis indicated that OmpW and TbpA proteins were highly conserved in different serotypes of swine Pasteurella multocida.SignalP 6.0 Server and TMHMM Server v.2.0 softwares predicted that OmpW protein had signal peptide and transmembrane region,while TbpA protein only had signal peptide without transmembrane region.ABCpred software analysis showed that OmpW and TbpA proteins had 14 and 22 B cell antigen epitopes,respectively.SOPMA and SWISS-MODEL softwares further predicted the protein structure of OmpW and TbpA,which showed that the alpha helix,beta turn,extended strand and random coil accounted for 16.18%,5.39%,36.76%,41.67%,and 40.42%,5.69%,16.77%,37.13%,respectively.The recombinant OmpW and TbpA proteins obtained by prokaryotic expression technique had strong antigenicity.Compared with control group,the serum specific antibody levels of OmpW and TbpA in serum were extremely significantly increased on 14 and 28 d after immunizing mice (P<0.01).The survival rate of mice in OmpW and TbpA groups was 50.0% and 37.5%,respectively.【Conclusion】 In this study,the OmpW and TbpA proteins of swine Pasteurella multocida were successfully expressed, the recombinant OmpW protein had strong immunogenicity and could be used as an effective candidate antigen for subunit vaccine against swine Pasteurella multocida.

Key words: swine Pasteurella multocida; OmpW; TbpA; expression; immunogenicity

CLC Number:

S852.61

ZHANG Zhewei, DAI Xiaotong, WEI Zudan, CHEN Yunpeng, GAN Jing, BEI Weicheng. Study on Prokaryotic Expression and Immunogenicity of OmpW and TbpA Proteins in Swine Pasteurella multocida[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1489-1498.

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Study on Prokaryotic Expression and Immunogenicity of OmpW and TbpA Proteins in Swine Pasteurella multocida

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