Zhang, Maojun, Meng, Fanliang, Cao, Fangfang, Qiao, Bo, Liu, Guodong, Liu, Hongying, Zhou, Yizhuang, Dong, Haiyan, Gu, Yixin, Xiao, Di, Zhang, Yongchan, and Zhang, Jianzhong
AbstractFourteen Campylobacter jejunigenes—porA, cadF, omp18, dnaK, flaC, peb1, peb2, peb3, peb4, ahpC, groEL, tuF, hipO,and Cj0069—were cloned and expressed in Escherichia coliBL21. The recombinant proteins were purified on histidine (His) and glutathione S–transferase (GST) trap columns using the ÄKTA Explorer 100 System. Recombinant proteins were visualized using sodium dodecyl sulfate–polyacrylamide gel electrophoresis and identified using matrix-assisted laser desorption ionization time-of-flight mass spectrometry. The antigenicities of these recombinant proteins were assessed by Western blotting and enzyme-linked immunosorbent assays with anti–C. jejuniimmune rabbit sera. Four recombinant proteins, including rGST-PorA, rHis-CadF, rGST-GroEL, and rGST-TuF, demonstrated reactions with both anti-serum and preimmune serum, while rHis-DnaK, rGST-FlaC, rGST-PEB2, rGST-PEB3, rGST-PEB4, and rGST-HipO showed variable antigenicity characteristics to the anti-sera derived from different C. jejunistrains. rHis-Omp18, rHis-PEB1, and rGST-AhpC demonstrated universal and specific antigenities with the entire anti-sera panel tested in this present study, while recombinant rGST-Cj0069 and rHis-DnaK did not react with any of the anti–C. jejunisera tested. In conclusion, rGST-AhpC may be useful as a potential serodiagnostic antigen for C. jejuniinfection.