Complementary DNA cloning and expression in Escherichia coli of Aln g I, the major allergen in pollen of alder (Alnus glutinosa).
Previous data showed that the major pollen allergens from trees of the order Fagales, in particular alder, birch, hazel, and hornbeam, are highly interrelated. As only the complete amino acid sequence of Bet v I, the major allergen from birch, has been known, it was of interest to obtain the primary structure of other major allergens of this group, to attribute IgE-binding properties to certain features of the amino acid sequences of those allergens. cDNA was synthesized from alder pollen mRNA, sequence-specifically amplified by polymerase chain reaction and cloned into plasmid bluescript. Comparison of the deduced amino acid sequences of Aln g I and Bet v I revealed a 86.8% homology. The Aln g I encoding cDNA was subcloned into pKK223-3 and expressed in Escherichia coli as a full-length nonfusion protein. The recombinant Aln g I bound IgE from tree pollen-allergic patients and was shown to share IgE-epitopes with Bet v I by inhibition studies with recombinant Bet v I. Computer-aided calculations predicted epitopes in both Aln g I and Bet v I at the same position; the Bet v I molecule was predicted to possess two additional epitopes near the N-terminus of the molecule.
Breiteneder
H
H
Institute of General and Experimental Pathology, University of Vienna, Austria.
Ferreira
F
F
Reikerstorfer
A
A
Duchene
M
M
Valenta
R
R
Hoffmann-Sommergruber
K
K
Ebner
C
C
Breitenbach
M
M
Kraft
D
D
Scheiner
O
O
D000485:Allergens / Q000737:chemistry*; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D001483:Base Sequence; D003001:Cloning, Molecular; D003201:Computers; D004247:DNA / Q000235:genetics*; D004926:Escherichia coli / Q000276:immunology*; D005784:Gene Amplification; D015151:Immunoblotting; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry*; D011058:Pollen / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D014197:Trees
D000485:Allergens; C067879:Aln g I protein, Alnus glutinosa; D052179:Antigens, Plant; D010940:Plant Proteins; D011994:Recombinant Proteins; C061312:Bet v 1 allergen, Betula; D004247:DNA