Cloning, expression, and antigenicity of 14 proteins from Campylobacter jejuni.

Maojun Zhang, Fanliang Meng, Fangfang Cao, Bo Qiao, Guodong Liu, Hongying Liu, Yizhuang Zhou, Haiyan Dong, Yixin Gu, Di Xiao, Yongchan Zhang, Jianzhong Zhang

National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China. zhangmaojun@icdc.cn

Foodborne pathogens and disease 2012 Aug

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Fourteen Campylobacter jejuni genes--porA, cadF, omp18, dnaK, flaC, peb1, peb2, peb3, peb4, ahpC, groEL, tuF, hipO, and Cj0069--were cloned and expressed in Escherichia coli BL21. 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. jejuni immune 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. jejuni strains. 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. jejuni sera tested. In conclusion, rGST-AhpC may be useful as a potential serodiagnostic antigen for C. jejuni infection.

Citation

Maojun Zhang, Fanliang Meng, Fangfang Cao, Bo Qiao, Guodong Liu, Hongying Liu, Yizhuang Zhou, Haiyan Dong, Yixin Gu, Di Xiao, Yongchan Zhang, Jianzhong Zhang. Cloning, expression, and antigenicity of 14 proteins from Campylobacter jejuni. Foodborne pathogens and disease. 2012 Aug;9(8):706-12