ELISA Expression and Identification of Recombinant A Domain of Adhesin Fnbp A from Staphylococcus aureus in Bovine Milk

Abstract

Abstract: Fibronectin binding protein A plays an important role in Staphylococcus aureus early infection, and is also a potential target of immunization. We subcloned the A domain of FnbpA into the prokaryotic expression vector pGEX and transformed it to the BL21 host cell. After removing the GST tag the purified purpose protein was analyzed by SDS-PAGE and approximately 63 kD exogenous protein was observed by SDS-PAGE. The purified protein without GST tag was emulsified by Freund’s adjuvant and injected rabbits subcutaneously to produce hyperimmune serum. The antibody titer and the binding capacity between the S. aureus and rabbit serum were tested by ELISA. The result indicated the antibody titer could reach 6.7×10⁶ and the antibody could recognize the antigen of S. aureus in vitro.

Key words: S. aureus, FnbpA, Prokaryotic expression, Protein purification, Immunogenicity ELISA

Su Yan, Wang Shimin, Li Ying, Wei Haina, Shao Jungao, Zhang Baojiang, Yin Tao. ELISA Expression and Identification of Recombinant A Domain of Adhesin Fnbp A from Staphylococcus aureus in Bovine Milk[J]. Biotechnology Bulletin, 2013, 0(4): 136-139.

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