Identification and stability of a 30-kDa nonstructural protein encoded by mRNA 2 of mouse hepatitis virus in infected cells.
A bacterial expression vector encoding a fusion protein containing almost the entire first open reading frame (ORF1) of mRNA 2 of MHV-A59 has been constructed. The purified fusion protein was used to raise antibodies to the protein encoded by mRNA 2 ORF1. Specificity of the antibodies was verified by immunoprecipitation of the in vitro translation product of ORF1, which was reconstructed downstream of a T7 promoter. In vivo the antiserum reacted specifically with a 30-kDa protein synthesized in MHV-A59- and MHV-JHM-infected cells. This 30-kDa protein could not be identified in purified virions and is therefore a nonstructural viral protein. The expression pattern of this 30-kDa nonstructural viral protein in infected cells was shown to be identical to that of the viral structural proteins. However, in comparison to the nucleocapsid protein pulse-chase studies revealed a relative short half life for this 30-kDa protein in vivo.
Bredenbeek
P J
PJ
Institute of Virology, Veterinary Faculty, State University of Utrecht, The Netherlands.
Noten
A F
AF
Horzinek
M C
MC
Spaan
W J
WJ
D000818:Animals; D002460:Cell Line; D002472:Cell Transformation, Viral; D003001:Cloning, Molecular; D005814:Genes, Viral; D008970:Molecular Weight; D006517:Murine hepatitis virus / Q000235:genetics*; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics*; D015183:Restriction Mapping; D014760:Viral Fusion Proteins / Q000032:analysis / Q000235:genetics*
D012333:RNA, Messenger; D014760:Viral Fusion Proteins