Cloning, expression,purification and immunological identification of Der f 11, paramyosin,a recombinant allergen of dust mite
10.3969/j.issn.1000-484X.2014.06.004
- VernacularTitle:粉尘螨重组过敏原Derf11（副肌球蛋白）克隆表达、纯化及免疫学鉴定
- Author:
Congli JIANG
;
Yulan WU
;
Peng XING
;
Pingchang YANG
;
Zhigang LIU
- Publication Type:Journal Article
- Keywords:
Dermatophagoides-farinae;
Recombination-Der-f-11;
Expression;
Western-blot;
Specific-IgE
- From:
Chinese Journal of Immunology
2014;(6):736-739
- CountryChina
- Language:Chinese
-
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.
