The integral membrane protein from a virulent isolate of transmissible gastroenteritis virus: molecular characterization, sequence and expression in Escherichia coli.
Subgenomic mRNA from a virulent isolate of porcine transmissible gastroenteritis virus (TGEV) was used to produce cDNA clones. Part of a new clone and a previously reported clone were sequenced and used to construct the viral gene for integral membrane protein. A single open reading frame (ORF) encoding a polypeptide of 262 amino acids, relative molecular mass (Mr) 29,459, was identified. The positive identification of the polypeptide as the integral membrane protein was demonstrated by the production in E. coli of a chimaeric protein comprising most of the ORF encoding the Mr 29,459 polypeptide and beta-galactosidase. The chimaeric protein reacted with a specific monoclonal antibody to viral integral membrane protein and antibodies raised against the chimaeric protein immune precipitated the viral protein. Comparison with the sequence of an avirulent isolate indicates amino acid residues that may be important in pathogenicity.
Britton
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Division of Microbiology, AFRC Institute for Animal Health, Compton Laboratory, Berkshire, UK.
Cármenes
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Garwes
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DJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D002213:Capsid / Q000235:genetics*; D003001:Cloning, Molecular; D003332:Coronaviridae / Q000235:genetics*; D004279:DNA, Viral; D004926:Escherichia coli / Q000235:genetics*; D005814:Genes, Viral; D008969:Molecular Sequence Data; D010957:Plasmids; D012367:RNA, Viral / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D005760:Transmissible gastroenteritis virus / Q000235:genetics* / Q000472:pathogenicity; D014774:Virulence
D004279:DNA, Viral; D012367:RNA, Viral; D011994:Recombinant Proteins