Cloning, expression,purification and immunological identification of Der f 11, paramyosin,a recombinant allergen of dust mite / 中国免疫学杂志

ABSTRACT

Objective: To clone , express and purify of Dermatophagoides farinae ( Der f 11 ) , and then test its immunogenicity.Methods:The gene coding for Der f 11 was synthesized ,and was then linked with the pET-32a vector.The expression plasmid pET32a(+)-Der f 11 was induced by IPTG.After purification of recombinant allergens Der 11 proteins through the Ni +affinity chromatography ,immunological allergic patients serum as the Primary antibody.Results: We obtain high purity recombinant Der f 11 protein.The results of SDS-PAGE show that the expression product is about 118 KD.Recombinant allergen Der f 11 test 15 dust mites allergic patients serum specific IgE , positive rate was 20%.Conclusion: Recombinant allergen Der f 11 obtained has the similar immunologic activity to natural Der f 11 protein.It can lay the foundation for the specific diagnosis ,treatment and further experimental studies of the dust mite allergy disease.