Expressions of recombinant venom allergen, antigen 5 of yellowjacket (Vespula vulgaris) and paper wasp (Polistes annularis), in bacteria or yeast.
Antigen 5 is a major allergen of vespid venom. It has partial sequence identity with proteins from diverse sources. The biologic function of Ag 5 and its related proteins is not known. We are interested in the expression of Ag 5 with the native conformation of the natural protein since its B cell epitopes are mainly of the discontinuous type. When expressed in bacteria, recombinant Ag 5 formed an insoluble intracellular product, and it did not translocate from cytoplasm to periplasm by the addition of a pelB leader sequence to the cloned protein. When expressed in yeast Pichia pastoris, Ag 5 was secreted because the cloned protein contained a yeast alpha signal leader sequence. Recombinant Ag 5 from yeast was shown to have the native structure of the natural protein and the recombinant Ag 5 from bacteria did not. This was shown by comparison of their solubility, electrophoretic behavior, disulfide bond content, CD spectrum, and binding of IgE antibodies from allergic patients and IgG antibodies from mice immunized with natural Ag 5 or recombinant Ag 5s from yeast or bacteria. These studies were made with Ag 5s from yellowjacket (Vespula vulgaris) and paper wasp (Polistes annularis).
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The Rockefeller University, 1230 York Avenue, New York, New York 10021-6399, USA.
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D000595:Amino Acid Sequence; D000818:Animals; D001419:Bacteria / Q000276:immunology*; D001483:Base Sequence; D002852:Chromatography, Ion Exchange; D002942:Circular Dichroism; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010843:Pichia / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014862:Wasp Venoms / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D014863:Wasps / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*
D011994:Recombinant Proteins; D014862:Wasp Venoms; C040583:white-faced hornet venom antigen 5