title: The construction of a cDNA expression library for the sheep scab mite Psoroptes ovis.
abstract: The need for alternative control strategies for sheep scab is critical. One approach is to develop vaccines based on 'concealed' antigens derived from Psoroptes ovis. This strategy requires the identification and characterisation of potential target antigens, which has been hampered by the problem of limited biological material for isolation of protein antigens. To aid the discovery of P. ovis antigens and to provide a resource for generating recombinant protein, we constructed a P. ovis cDNA expression library, using total RNA isolated from 250 mg of mixed-stage P. ovis and the Clontech SMART cDNA synthesis kit. The presence of P. ovis-specific sequences was confirmed using PCR amplification and sequencing of actin. The sequences of cDNA inserts from six random clones included one with high homology to the Dermatophagoides pteronyssinus (house dust mite) antigen p Dp15. This is a glutathione S-transferase known to be an important house dust mite antigen. We conclude that this library will be a useful tool for the identification of potential target antigens for the immunological control of P. ovis and to further our understanding of the pathology of sheep scab.
authors.affiliation: School of Biology, The University of Leeds, UK.
authors.forename: A J
authors.initials: AJ
authors.lastname: Lee
authors.forename: R E
authors.initials: RE
authors.lastname: Isaac
authors.forename: D
authors.initials: D
authors.lastname: Coates
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000737:chemistry; D015723:Gene Library; D008924:Mite Infestations / Q000276:immunology / Q000517:prevention & control / Q000662:veterinary*; D008925:Mites / Q000737:chemistry*; D008969:Molecular Sequence Data; D010802:Phylogeny; D010957:Plasmids / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012313:RNA / Q000302:isolation & purification; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012756:Sheep; D012757:Sheep Diseases / Q000276:immunology / Q000469:parasitology*; D013983:Tick Control / Q000379:methods*
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D012313:RNA