cDNA sequences encoding secretory proteins from a library constructed using mRNA isolated from C.pulicaris salivary glands.
Female Culicoides (midges) were caught by suction trap as they alighted for feed on a horse.  Those with wing patterning of C.pulicaris group midges (consisting of C. pulicaris and C. punctatus spp.) were separated and their salivary glands dissected.  mRNA isolated from these tissues was used to construct a cDNA library using the Clontech SMART cDNA synthesis kit.  One hundred and fifty clones from this library were sequenced using conventional sanger sequencing.  Those with high quality sequence data representing a complete open reading frame including a secretory signal sequence through to the polyA tail were selected for further analysis and expression studies to assess their role as allergens causing type I hypersensitivity in horses.
Female Culicoides (midges) were caught by suction trap as they alighted for feed on a horse.  Those with wing patterning of C.pulicaris group midges (consisting of C. pulicaris and C. punctatus spp.) were separated and their salivary glands dissected.  mRNA isolated from these tissues was used to construct a cDNA library using the Clontech SMART cDNA synthesis kit.  One hundred and fifty clones from this library were sequenced using conventional sanger sequencing.  Those with high quality sequence data representing a complete open reading frame including a secretory signal sequence through to the polyA tail were selected for further analysis and expression studies to assess their role as allergens causing type I hypersensitivity in horses.
University of Bristol