Premium
Nucleotide sequence of cDNA encoding the group II allergen of cocksfoot/orchard grass ( Dactylis glomerata), Dac g II
Author(s) -
Roberts A. M.,
Bevan L. J.,
Flora P. S.,
Jepson I.,
Walker M. R.
Publication year - 1993
Publication title -
allergy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.363
H-Index - 173
eISSN - 1398-9995
pISSN - 0105-4538
DOI - 10.1111/j.1398-9995.1993.tb00758.x
Subject(s) - dactylis glomerata , biology , complementary dna , microbiology and biotechnology , nucleic acid sequence , immunoscreening , fusion protein , lolium perenne , peptide sequence , gene , genetics , cdna library , botany , recombinant dna , poaceae
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 β‐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 λgt 11 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.