High-level expression of tree pollen isoallergens in Escherichia coli.
cDNAs coding for the major allergen of alder (Alnus glutinosa) pollen Aln g 1, for nine isoforms of Bet v 1, the major birch (Betula verrucosa) pollen allergen, and for four isoforms of Cor a 1, the major allergen of hazel (Corylus avellana) pollen, were inserted into the plasmid pMW175 or pMW 172 and expressed in Escherichia coli as recombinant non-fusion proteins. These constructs produced between 20 and 160 mg protein/l. The recombinant tree pollen isoallergens were tested in immunoblots for their antibody binding properties. For this purpose, we used two monoclonal antibodies (BIP 1 and BIP 4) raised against natural Bet v 1, a polyclonal rabbit anti-recombinant Bet v 1a, as well as serum IgE from allergic patients. Our results show that this expression system is suitable for the production of milligram amounts of tree pollen isoallergens which can be used for the characterization of allergenic epitopes recognized by T and B cells.
Weiss
C
C
Institut für Genetik und Allgemeine Biologie, Universität Salzburg, Austria.
Kramer
B
B
Ebner
C
C
Susani
M
M
Briza
P
P
Hoffmann-Sommergruber
K
K
Breiteneder
H
H
Kraft
D
D
Scheiner
O
O
Breitenbach
M
M
Ferreira
F
F
D000485:Allergens / Q000096:biosynthesis / Q000235:genetics*; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics* / Q000276:immunology; D005822:Genetic Vectors / Q000276:immunology*; D015151:Immunoblotting; D008969:Molecular Sequence Data; D011058:Pollen / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000276:immunology*; D014197:Trees / Q000276:immunology
D000485:Allergens; D011994:Recombinant Proteins