Detection of Mouse Hepatitis Virus Nonstructural Proteins Using Antisera Directed Against Bacterial Viral Fusion Proteins
Mouse hepatitis virus, strain A59 cDNAs were inserted into the procaryotic fusion vector pGE374. RecA/viral/LacZ tripartite fusion proteins were synthesized from these plasmids and purified from E. coli. Antisera were raised in rabbits against these fusion proteins. Viral nonstructural proteins were detected in infected murine fibroblasts and glial cells. The anti-gene B, 0RF1 sera detect a 3OK cytoplasmic protein while the anti-gene E, ORF2 sera detect a 9.6K protein. Sera raised against proteins encoded in cDNAs from 5′ portions of gene A immunoprecipitate the 200–250K polypeptides synthesized in vitro from genome RNA. Antisera raised against proteins encoded in both 5′ and 3′ portions of gene A immunoprecipitate membrane associated polypeptides of 150K and >600K from MHV infected cells.
KeywordsHepatitis Codon Recombination Influenza Fractionation
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