Protein Coding Assignment for the Genome of Epizootic Haemorrhagic Disease Virus

Abstract

SUMMARYViral genomic RNA was purified from BHK-21 cells infected with epizootic haemorrhagic disease virus and the 10 dsRNA genome segments were isolated by polyacrylamide gel electrophoresis. These genome segments were translated in vitro using the rabbit reticulocyte lysate system and the synthesized proteins were detected by immune precipitation and gel electrophoresis. This allowed the assignment of protein coding to the genome segments and the identification of two additional virusspecified proteins not readily detectable in lysates of virus-infected cells.

Sequence of genome segment 9 of bluetongue virus (serotype 1, South Africa) and expression analysis demonstrating that different forms of VP6 are derived from initiation of protein synthesis at two distinct sites

“…Recently published data on the genome coding assignment of two other members of the orbivirus genus, African horsesickness virus (Grubman & Lewis, 1992) and epizootic haemorrhagic disease virus (Mecham & Dean, 1988), indicate that a protein doublet is encoded by $9 of these two isolates. A protein doublet is also apparent after SDS-PAGE of the in vitro translation products from AHSV-9 $9 (R. O'Hara & P. P. C. Mertens, unpublished results).…”

A Multiplex Real-Time Reverse Transcription Polymerase Chain Reaction Assay for Detection and Differentiation of Bluetongue Virus and Epizootic Hemorrhagic Disease Virus Serogroups

“…The protein coding assignments for BTV and EHDV are identical, consisting of 10 genome segments that encode 7 structural and 4 nonstructural proteins. 20 Developing molecular diagnostics is complicated by the multiple serotypes of these viruses: 24 for BTV and 8 for EHDV. 19 The outer capsid protein, VP2, is the primary serotype determinant, but the second outer capsid protein VP5 can also influence serotypic epitopes.…”

A comparison of the genes which encode non-structural protein NS3 of different orbiviruses

“…AHSV and EHDV have seven structural proteins each (Bremer, 1976;Huismans et al, 1979;Mecham & Dean, 1988), whereas nine were identified for a member of the Palyam serogroup (Whistler & Swanepoel, 1990). The nonstructural proteins (NS1, NS2 and NS3) of EHDV were identified by in vitro translation (Mecham & Dean, 1988), and a comparison of virus-infected cell lysates with purified viral proteins revealed at least three nonstructural Palyam virus proteins, designated NS1, NS2a and NS2b (Whistler & Swanepoel, 1990).…”