Detection of a murine coronavirus nonstructural protein encoded in a downstream open reading frame

• Published in: Virology (New York, N.Y.) Vol. 164; no. 1; pp. 156 - 164
• Main Authors: Leibowitz, Julian L., Perlman, Stanley, Weinstock, George, Devries, James R., Budzilowicz, Carol, Weissemann, Jane M., Weiss, Susan R.
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 01.05.1988; Elsevier; Published by Elsevier Inc
• Subjects: Biological and medical sciences; Escherichia coli; Fundamental and applied biological sciences. Psychology; Genes; Genes, Viral; Genetic Vectors; Genetics; Microbiology; Murine hepatitis virus - genetics; Plasmids; Recombinant Fusion Proteins - genetics; Viral Proteins - genetics; Virology; Virus; Plasmid; Proteins; Immunocytochemistry; Infected cell; Coronaviridae; Gene expression; Hybrid gene; Murine hepatitis virus; Immunological method
• ISSN: 0042-6822; 1096-0341
• DOI: 10.1016/0042-6822(88)90631-9

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 β-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.