cDNA sequences encoding secretory proteins from a library constructed using mRNA isolated from C.obsoletus salivary glands and from an assembly created following Ion Torrent sequencing of C.obsoletus cDNA.
Female Culicoides (midges) were caught by suction trap as they alighted for feed on a horse.  Those with wing patterning of C.obsoletus group midges 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. In addition the cDNA was sequenced by Ion Torrent sequencing and Trinity assembly, data are available at the University of Bristol data repository, data.bris, at https://doi.org/10.5523/bris.2z11o8xv6g9j62w5gtuafpenst.  The assembly was searched for sequences with similarity to all known salivary gland secretory proteins from Culicoides spp. and the results combined with those from the cDNA library to provide a more comprehensive list of C.obsoletus salivary proteins.
Female Culicoides (midges) were caught by suction trap as they alighted for feed on a horse.  Those with wing patterning of C.obsoletus group midges 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. In addition the cDNA was sequenced by Ion Torrent sequencing and Trinity assembly, data are available at the University of Bristol data repository, data.bris, at https://doi.org/10.5523/bris.2z11o8xv6g9j62w5gtuafpenst.  The assembly was searched for sequences with similarity to all known salivary gland secretory proteins from Culicoides spp. and the results combined with those from the cDNA library to provide a more comprehensive list of C.obsoletus salivary proteins.
University of Bristol