China Animal Husbandry and Veterinary Medicine ›› 2025, Vol. 52 ›› Issue (2): 801-810.doi: 10.16431/j.cnki.1671-7236.2025.02.030

• Preventive Veterinary Medicine • Previous Articles    

Bioinformatics Analysis and Prokaryotic Expression of EgBAL Protein of Echinococcus granulosus

ZHAO Wenqing1,2, BO Xinwen1,2,3, PU Na2,3, ZHANG Yuxia2,3, CHEN Xuke2,3, ZHANG Yanyan2, SUN Yan2, QI Meng1,2, WANG Zhengrong2   

  1. 1. College of Animal Science and Technology, Tarim University, Alar 843300, China;
    2. Provincial-Ministry Joint State Key Laboratory of Sheep Genetic Improvement and Healthy Breeding, Institute of Animal Husbandry and Veterinary Medicine, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832000, China;
    3. College of Animal Science and Technology, Shihezi University, Shihezi 832000, China
  • Revised:2024-08-07 Published:2025-01-24

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Abstract: 【Objective】 The aim of this study was to explore the bioinformatics characteristics of bile salt activated lipase (EgBAL) in Echinococcus granulosus and perform prokaryotic expression to detect the immunogenicity of EgBAL recombinant protein,provide theoretical basis for antigen screening of cystic echinococcosis vaccine. 【Method】 EgBAL gene sequence (GenBank accession No.:HM748917) of Echinococcus granulosus was obtained from NCBI database,specific primers were designed,and EgBAL gene was amplified and cloned by PCR using the cDNA of Ecanococcus granulosus as template.Phylogenetic tree was constructed by NJ method using Mega 7.0 software.The physical and chemical properties and structural characteristics of EgBAL protein were predicted and analyzed by bioinformatics softwares.Recombinant pET-22b plasmid containing EgBAL gene was constructed and transformed into Escherichia coli BL21(DE3) competent cells to induce expression of EgBAL recombinant protein.The protein expression was detected by SDS-PAGE and purified.The immunogenicity of the recombinant protein was analyzed by Western blotting. 【Result】 The fragment size of the EgBAL gene in Echinococcus granulosus was 1 020 bp.The phylogenetic tree showed that the EgBAL protein was in the same branch as the EgBAL protein of the Tupaia belangeri with a close evolutionary distance and a similarity as high as 99.60%.Bioinformatics analysis revealed that the EgBAL gene was 1 014 bp in length,encoding 337 amino acids.The molecular weight of EgBAL protein was 37 089.59 u,with a theoretical isoelectric point of 4.77,an instability index of 44.72,indicating an unstable protein,a fat coefficient of 66.11,a hydrophilicity of －0.429,a hydrophilic protein with no transmembrane region or signal peptide.EgBAL protein had 14 potential B-cell epitopes,and the secondary structure included alpha helix,extended chain,beta turn and random coil,accounting for 27.90%,12.76%,12.76% and 56.08%,respectively.In addition,EgBAL protein contained 35 phosphorylation sites,including 12 serine phosphorylation sites,16 threonine phosphorylation sites and 7 tyrosine phosphorylation sites.In this study,the recombinant pET-22b-EgBAL plasmid was successfully constructed,and the size of EgBAL recombinant protein was about 38 ku.Western blotting results showed that the recombinant protein could be recognized by the positive sera of Kunming mice and dogs infected with Echinococcus granulosus,and had good immunogenicity. 【Conclusion】 This study successfully cloned the EgBAL gene of Echinococcus granulosus,expressed the recombinant EgBAL protein,and preliminarily confirmed the recombinant EgBAL protein had good immunogenicity,suggesting its potential as a vaccine or diagnostic candidate antigen for cystic echinococcosis.These findings provided a foundational scientific basis for further investigation into the functional role of the EgBAL protein.

Key words: Echinococcus granulosus; EgBAL gene; bioinformatics analysis; prokaryotic expression

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