Detection of a murine coronavirus nonstructural protein encoded in a downstream open reading frame.
Mouse hepatitis virus (MHV) gene 5 contains two open reading frames. We have expressed the second open reading frame of this gene (gene 5 ORF 2) in an Escherichia coli expression system. This system utilized a plasmid which contained the promoter and the first 36 codons of the recA gene fused in frame with the MHV gene 5 ORF 2, which is fused in turn to the beta-galactosidase gene. The protein product of this gene fusion was used to raise antibody to gene 5 ORF 2. The specificity of the antibody was verified by immunoprecipitation of the in vitro transcribed and translated protein product of gene 5 ORF 2. The second reading frame of MHV gene 5 was shown to be expressed during the course of infection by immunocytochemistry and radioimmunoprecipitation using the antibody raised against the E. coli fusion protein and by two-dimensional gel electrophoresis.
Leibowitz
J L
JL
Department of Pathology and Laboratory Medicine, University of Texas Health Science Center, Houston 77225.
Perlman
S
S
Weinstock
G
G
DeVries
J R
JR
Budzilowicz
C
C
Weissemann
J M
JM
Weiss
S R
SR
D005796:Genes; D005814:Genes, Viral; D005822:Genetic Vectors; D006517:Murine hepatitis virus / Q000235:genetics*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics*
D011993:Recombinant Fusion Proteins; D014764:Viral Proteins