Expression of Dermatophagoides pteronyssinus allergen, Der p II, in Escherichia coli and the binding studies with human IgE.
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 beta-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.
Clinical Immunology Research Unit, Princess Margaret Hospital, Subiaco, Australia.
K Y
KY
Chua
R J
RJ
Dilworth
W R
WR
Thomas
D000485:Allergens / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D000818:Animals; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology*; D008925:Mites / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology
D000485:Allergens; D011994:Recombinant Proteins; D007073:Immunoglobulin E