Nucleotide sequence of cDNA encoding the group II allergen of cocksfoot/orchard grass (Dactylis glomerata), Dac g II.
Cocksfoot/orchard grass (Dactylis glomerata) anther cDNA clones encoding the group II allergen Dac g II were previously isolated on the basis of immunoreactivity of human, rabbit, and murine antibodies with a 24-kDa protein expressed as a fusion protein with beta-galactosidase. Nucleotide sequencing reveals an open reading frame predicting expression of a 98-amino-acid (11-kDa) polypeptide exhibiting > 90% homology with the group II allergen of Lolium perenne, Lol p II. In vitro translation of different sized clone fragments generated by polymerase chain amplification confirms eukaryotic expression of a 10-12-kDa polypeptide by SDS-PAGE and the position of a translational stop apparently unrecognized during expression of lambda gt11 in E. coli. The unusual characteristics of the prokaryote-expressed fusion proteins may be exerting conformational alterations in Dac g II, as reflected by previous demonstrations of differences in human IgE immunoreactivity. Northern blot analysis using PCR-generated partial and full-length probes suggests that group II allergens may be encoded by a different family or families of temporally expressed genes from those encoding group I major allergens, although a group I gene may have been the progenitor.
Roberts
A M
AM
University Department of Clinical Biochemistry, Wolfson Research Laboratories, Queen Elizabeth Medical Centre, Birmingham, UK.
Bevan
L J
LJ
Flora
P S
PS
Jepson
I
I
Walker
M R
MR
D000485:Allergens / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D017343:Genes, Plant; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics / Q000276:immunology; D006109:Poaceae / Q000235:genetics / Q000276:immunology; D011058:Pollen / Q000235:genetics* / Q000276:immunology*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics
D000485:Allergens; D017931:DNA Primers; D018076:DNA, Complementary; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins