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Updated: May 20, 2026
Detection and Isolation of Campylobacter spp. from Raw Meat
Published on: February 23, 2024
Cloning, expression, and antigenicity of 14 proteins from Campylobacter jejuni
Maojun Zhang1, Fanliang Meng, Fangfang Cao
1National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China. zhangmaojun@icdc.cn
Insights
Researchers explored fourteen Campylobacter jejuni genes to identify potential antigens for diagnosing infections. Recombinant glutathione S-transferase-AhpC (rGST-AhpC) showed promising reactivity, suggesting its utility in serodiagnostic tests for Campylobacter jejuni.
Area of Science:
- Microbiology and Immunology
- Bacterial Pathogenesis
- Recombinant Protein Expression
Background:
- Campylobacter jejuni is a leading cause of bacterial gastroenteritis worldwide.
- Accurate and rapid diagnostic tools are crucial for managing C. jejuni infections.
- Identifying specific C. jejuni antigens is key for developing effective serodiagnostic assays.
Purpose of the Study:
- To clone, express, and purify fourteen selected C. jejuni genes in Escherichia coli.
- To evaluate the antigenicity of the resulting recombinant proteins using immune sera.
- To identify potential C. jejuni serodiagnostic antigens.
Main Methods:
- Cloning and expression of fourteen C. jejuni genes in E. coli BL21.
- Purification of recombinant proteins using histidine (His) and glutathione S-transferase (GST) affinity chromatography.
- Assessment of protein identity and antigenicity via SDS-PAGE, mass spectrometry, Western blotting, and ELISA using anti-C. jejuni sera.
Main Results:
- Recombinant proteins were successfully expressed and purified.
- rHis-Omp18, rHis-PEB1, and rGST-AhpC exhibited universal and specific antigenicity against the tested sera.
- Recombinant rGST-Cj0069 and rHis-DnaK did not react with any anti-C. jejuni sera.
Conclusions:
- Several C. jejuni recombinant proteins demonstrated antigenicity, with varying specificities.
- Recombinant glutathione S-transferase-AhpC (rGST-AhpC) showed significant promise as a potential serodiagnostic antigen.
- Further validation of rGST-AhpC could lead to improved diagnostic methods for C. jejuni infections.
Abstract:
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.
