Recombinant Expression, Purification and Identification of Egg Yolk Allergen Gal d 6

Objective:To establish an egg yolk Gal d 6 expressed engineering strain and identify the immunological activity of rGal d 6 protein. Methods:The Gal d 6 gene was directly synthesized, and then cloned into the vector pET-28a. The constructed recombinant plasmid was transformed into E.coli BL21 and the protein was induced expression by IPTG and purified by Ni-NTA.Identification of rGal d 6 immunoreactivity was detected by indirected ELISA and western blot. Results:The optimum expression conditions for the engineered bacteria were 37℃, 0.5 mmol/L IPTG, 2 hours, and the yield could reach 9.1 mg per liter. A single band was obtained by Ni-NTA. The recombinant protein had good reactivity with egg allergy serum. Conclusion:An engineered strain with stable expression of Gal d 6 protein with immunoreactivity was established.