title: Molecular characterization, expression and localization of tropomyosin and paramyosin immunodominant allergens from sheep scab mites (Psoroptes ovis).
abstract: cDNAs encoding the immunodominant allergens tropomyosin and paramyosin were amplified from RNA extracted from the sheep scab mite Psoroptes ovis. The tropomyosin cDNA contained an open reading frame (ORF) of 852 bp which encoded a predicted protein with 98% and 97% identity to the house dust mite allergens Der f 10 and Der p 10 respectively. The complete paramyosin ORF generated by RT-PCR was 2625 bp in length and encoded an 875aa predicted protein of 102.6 kDa with 97%, 95% and 89% identity to the paramyosins of Dermatophagoides pteronyssinus (Der p 11), Sarcoptes scabiei and Blomia tropicalis (Blo t 11) respectively. Full length tropomyosin and truncated and full-length paramyosin were expressed as recombinant proteins. IgG and IgE in sera from sheep with a 6-week duration primary infestation of P. ovis did not detect either full-length or truncated recombinant paramyosin. IgG in both infested and naïve sheep sera detected recombinant tropomyosin, suggesting cross-reactivity to tropomyosin and to other invertebrate species to which the sheep may have been exposed. Staining with antibodies directed against tropomyosin and paramyosin was observed throughout sections of P. ovis. Staining was especially prevalent in the anterior sections of the mites, possibly associated with locomotory muscles in this region.
authors.affiliation: Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, EH26 0PZ, Scotland. Alasdair.Nisbet@moredun.ac.uk
authors.forename: A J
authors.initials: AJ
authors.lastname: Nisbet
authors.forename: A
authors.initials: A
authors.lastname: MacKellar
authors.forename: H W
authors.initials: HW
authors.lastname: Wright
authors.forename: G P
authors.initials: GP
authors.lastname: Brennan
authors.forename: K Y
authors.initials: KY
authors.lastname: Chua
authors.forename: N
authors.initials: N
authors.lastname: Cheong
authors.forename: J E
authors.initials: JE
authors.lastname: Thomas
authors.forename: J F
authors.initials: JF
authors.lastname: Huntley
mesh_terms: D000485:Allergens / Q000276:immunology / Q000302:isolation & purification*; D000818:Animals; D001483:Base Sequence; D003429:Cross Reactions; D018076:DNA, Complementary / Q000737:chemistry; D016056:Immunodominant Epitopes / Q000276:immunology / Q000302:isolation & purification*; D008924:Mite Infestations / Q000276:immunology / Q000469:parasitology / Q000662:veterinary*; D008970:Molecular Weight; D016366:Open Reading Frames; D040042:Psoroptidae / Q000737:chemistry* / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012756:Sheep; D012757:Sheep Diseases / Q000276:immunology / Q000469:parasitology*; D014335:Tropomyosin / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000485:Allergens; D018076:DNA, Complementary; D016056:Immunodominant Epitopes; D014335:Tropomyosin