Recombinant Expression and Allergenicity Identification of Sesame Allergen Ses i 3

Abstract

Total RNA was extracted from sesame seeds, and the sesame Ses i 3 gene was obtained by using reverse transcription-polymerase chain reaction (RT-PCR). The plasmid pET-22b(+) was used to construct an expression vector, and the recombinant plasmid was transformed into competent Escherichia coli BL21 (DE3) host cells for induced expression. The expressed protein was purified by Ni2+-affinity chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that the molecular mass of the expressed protein was approximately 66 kDa. The ability of the recombinant and native Ses i 3 protein to induce sesame allergy in mice was compared by using a BALB/c mouse allergy model, which was found to be similar by measuring specific antibodies (immunoglobulin E (IgE), IgG1, and IgG2a), cytokines (interleukin (IL)-4, IL-5, interferon-γ (IFN-γ) and histamines). Hence, the recombinant Ses i 3 protein can be used for future studies on the allergenicity of sesame.

Keywords

• sesame allergen; ses i 3; recombinant protein; allergenicity; balb/c mice