现代牧业2025，Vol.9Issue(2)：24-32,9.
绵羊肺炎支原体黏附素P120571aa～703aa蛋白原核表达及免疫原性分析
Prokaryotic expression and immunogenicity analysis of adhesive P120571aa～703aa in Mycoplasma ovipneumoniae
摘要
Abstract
To identify candidate antigens for the development of a subunit vaccine and serological diagnosis against sheep contagious pleuropneumonia,this study employed bioinformatics tools IEDB(Immune Epitope Database)and ABCpred to predict B-cell epitopes on the P120 protein of Mycoplasma ovipneumoniae strain NM2010.Based on the bioinformatics predictions,the gene fragment encoding the dominant antigenic epitope-rich region(residues 540-710)of P120 was chemically synthesized and cloned into the expression vector pET-28a(+).The recombinant protein was expressed in E.coli,purified,and analyzed via Western blot.Rabbits were immunized with the recombinant protein to evaluate immune responses.ELISA was used to detect serum-specific IgG antibodies and cytokines(IL-2,IL-4,IL-10,IFN-γ).Results demonstrated that residues 571-703 of the P120 protein constitute a dominant antigenic epitope region.The synthetic gene(414 bp)corresponding to residues 571-703 was correctly amplified,and the expressed recombinant protein(P120571-703aa)exhibited a molecular weight of 17.2 kDa,primarily forming inclusion bodies.Western blot analysis confirmed its specificity against hyperimmune rabbit serum against Mycoplasma ovipneumoniae strain NM2010.Immunized rabbits developed high-titer IgG antibodies(1∶128,000),with significant increases in Th1/Th2 cytokines(P<0.01)compared to the control group.These findings indicate that the P120571-703aa recombinant protein effectively induces both humoral and cellular immunity,highlighting its potential as a candidate antigen for mycoplasmal pneumonia vaccines in sheep.关键词
绵羊肺炎支原体/黏附素P120/抗原表位/原核表达/反应原性/免疫原性Key words
Mycoplasma ovipneumoniae/adhesive P120/antigenic epitope/P120/pro-karyotic expression/antigenicity/immunogenicity分类
农业科技引用本文复制引用
徐春光,杨立霞,于宝莉,李艳,张月梅..绵羊肺炎支原体黏附素P120571aa～703aa蛋白原核表达及免疫原性分析[J].现代牧业,2025,9(2):24-32,9.基金项目
内蒙古自治区自然科学基金项目(2022MS03069). （2022MS03069）
