Expression of Dermatophagoides pteronyssinus Allergen, Der p II, in Escherichia coli and the Binding Studies with Human IgE
- 1 January 1990
- journal article
- research article
- Published by S. Karger AG in International Archives of Allergy and Immunology
- Vol. 91 (2) , 124-129
- https://doi.org/10.1159/000235102
Abstract
Lambda gt11 clones expressing the major house dust mite allergen, Der p II, have been reported to react with IgE in the serum of a high proportion of allergic patients. The clones described, however, only produced small quantities of protein which was not fused to the β-galactosidase of the vector. A construct of the Der p II is described which produces a fusion of Der p II, minus its leader sequence, with the glutathione-S transferase in the pGEX vector. This could be readily isolated and was shown to react with IgE in 22 of 24 patient sera showing reactivity to native Der p II, the sera not reacting having low reactivity to native protein. Absorption analysis showed that the recombinant material removed most of the IgE reactivity of patients to native Der p II. The construct described, therefore, should be valuable for quantitative studies of a pure mite allergen.Keywords
This publication has 4 references indexed in Scilit:
- Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferaseGene, 1988
- Epitope mapping of two major inhalant allergens, Der p I and Der f I, from mites of the genus Dermatophagoides.The Journal of Immunology, 1987
- Antigens and allergens from the common house dust mite Dermatophagoides pteronyssinusJournal of Allergy and Clinical Immunology, 1984
- “Western Blotting”: Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein AAnalytical Biochemistry, 1981