Expression and characterisation of the ovine respiratory syncytial virus (ORSV) G protein for use as a diagnostic reagent.
Respiratory syncytial virus (RSV) causes severe lower respiratory tract infection in children and calves. Antibodies to ovine RSV (ORSV) are common in sheep, but the clinical disease is not well defined. There is no report of ORSV infection in Australian sheep although respiratory distress syndrome has been described. This discrepancy may be due to the lack of a suitable diagnostic test. In this report, we have characterised the ORSV G protein in an attempt to study its relatedness to human and bovine RSV (HRSV, BRSV) and for use in the development of a suitable diagnostic assay. Full length and a truncated variant of ORSV G protein were expressed in mammalian cells and the expressed proteins characterised by indirect immunofluorescence and radioimmunoprecipitation assays. Our results indicate that like HRSV, the ORSV G protein is heavily glycosylated. The expressed protein was membrane bound as well as secreted and could be purified from culture supernatants and may be suitable for use in development of a diagnostic assay.
Ghildyal
R
R
Children's Virology Research Unit, Macfarlane Burnet Centre for Medical Research, Fairfield, Victoria, Australia.
Chapman
A
A
Peroulis
I
I
Mills
J
J
Meanger
J
J
D000818:Animals; D001315:Australia; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D002648:Child; D003001:Cloning, Molecular; D003937:Diagnosis, Differential; D019084:Fluorescent Antibody Technique, Indirect; D015707:HN Protein; D006801:Humans; D008969:Molecular Sequence Data; D011863:Radioimmunoassay; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis / Q000662:veterinary*; D018112:Respiratory Syncytial Virus, Bovine; D018113:Respiratory Syncytial Virus, Human; D012136:Respiratory Syncytial Viruses / Q000235:genetics / Q000302:isolation & purification*; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis* / Q000821:virology; D014162:Transfection; D014759:Viral Envelope Proteins; D014764:Viral Proteins / Q000032:analysis / Q000235:genetics
D015707:HN Protein; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C116204:attachment protein G