迟缓爱德华菌分子伴侣蛋白GroEL原核表达及免疫特性

Prokaryotic expression and immunological protection analysis of molecular chaperone protein GroEL of Edwardsiella.tarda

GroEL是细菌内分子伴侣蛋白,参与细菌多种生命活动,在布鲁菌、链球菌研究中,GroEL蛋白展示了良好的疫苗潜力。前期利用迟缓爱德华菌抗体Pull-down技术筛选到GroEL蛋白,揭示了该蛋白的潜在应用价值,本试验以迟缓爱德华菌GroEL蛋白作为研究对象,利用大肠杆菌表达系统表达纯化GroEL重组蛋白(rGroEL),进一步通过动物模型评估rGroEL的免疫效果。结果显示,rGroEL能够激发小鼠产生高水平的特异性抗体,具有较好的免疫原性,免疫小鼠对迟缓爱德华菌感染具有较好的抵抗力,相对保护率达95%。斑马鱼感染试验结果表明,rGroEL免疫对迟缓爱德华菌感染具有一定保护效果,保护率为60%。用rGroEL免疫斑马鱼可以产生针对嗜水气单胞菌的免疫保护力,保护率可达45%。本试验初步探索了迟缓爱德华菌重组蛋白rGroEL的免疫特性,为研制迟缓爱德华菌亚单位疫苗和核酸疫苗提供了依据。

GroEL,a molecular chaperone protein,plays importmant roles in vital movement of bacteria.This protein,when used as immunogen,displays excellent potential as vaccine in publication about Brucella and Streptococcus.In previous study,GroEL protein was determined to be a dominant antigen of Edwardsiella tarda(E.tarda)with antibody-based pull-down assay.Here,we performed expression and purification of recombinant GroEL protein(rGroEL)of E.tarda and further assessed the immunological characteristics of rGroEL.The data showed the recombinant protein stimulated high-level specific antibody and excellent protection against E.tarda infection in mice with 95%relative percent survival(RPS).Furthermore,the zebra fish received rGroEL injection displayed considerable resistance to E.tardainfection with RPS at 60%.Moreover,the zebra fish received rGroEL injection also showed a certain resistance to Aeromonas hydrophila infection with RPS at 45%.In sum,these work indicated the immunological effects and vaccine potential of rGroEL,which provide guidance and foundation for development of subunit vaccine and nucleic acid vaccine of E.tarda.