Eukaryotic expression of recombinant Pso o 1, an allergen from Psoroptes ovis, and its localization in the mite.
A cDNA encoding the immunogen Pso o 1 from Psoroptes ovis was obtained by polymerase chain reaction (PCR) amplification. The amplicon contained the entire coding sequence for the prepro-enzyme in an open reading frame (ORF) of 966 bp. This gene encoded a predicted protein of 322 amino acids (aa) with 64% aa identity (80% similarity) to the major house dust mite faecal allergen Der f 1. The pro-enzyme form of Pso o 1 was expressed as a recombinant protein in the Pichia pastoris-eukaryotic expression system. Maturation of the recombinant pro-enzyme by autocatalytic activation was not observed, and such maturation could not be achieved using a number of techniques known to activate recombinant Der p 1 and Der f 1 expressed in the same system. Serum raised against recombinant Pso o 1 cross-reacted with mature Der p 1 and allowed Pso o 1 to be immunolocalized to the gut of P. ovis.
Nisbet
A J
AJ
Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, EH26 0PZ, Scotland. Alasdair.Nisbet@moredun.ac.uk
MacKellar
A
A
McLean
K
K
Brennan
G P
GP
Huntley
J F
JF
D000485:Allergens / Q000032:analysis* / Q000235:genetics* / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D039741:Antigens, Dermatophagoides / Q000235:genetics; D060829:Arthropod Proteins; D001483:Base Sequence; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases; D008924:Mite Infestations / Q000276:immunology / Q000662:veterinary; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics; D040042:Psoroptidae / Q000033:anatomy & histology / Q000737:chemistry / Q000276:immunology*; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology; D012756:Sheep / Q000276:immunology; D012757:Sheep Diseases / Q000276:immunology
D000485:Allergens; D039741:Antigens, Dermatophagoides; D060829:Arthropod Proteins; D011994:Recombinant Proteins; D003546:Cysteine Endopeptidases; C451159:Dermatophagoides farinae antigen f 1; C451162:Dermatophagoides pteronyssinus antigen p 1