Sequence of the nucleoprotein gene from a virulent British field isolate of transmissible gastroenteritis virus and its expression in Saccharomyces cerevisiae

Mol Microbiol. 1988 Jan;2(1):89-99.

Abstract

Subgenomic mRNA from a virulent isolate of porcine transmissible gastroenteritis virus (TGEV) was used to produce cDNA which was sequenced. Two non-overlapping open reading frames (ORFs) were identified. The largest, encoding a polypeptide of 382 amino acids (relative molecular mass (Mr) 43,483), was shown to be the viral nucleoprotein gene. The second ORF, found 3' to the larger ORF, encodes a polypeptide of 78 amino acids (Mr 9068) which has yet to be assigned to a viral product. The nucleoprotein gene was expressed in yeast cells under the control of two types of yeast promoters: the constitutive PGK promoter, and the inducible GAL1 promoter. Yeast cells containing recombinant plasmids, with the nucleoprotein gene in the correct orientation, produced a polypeptide of Mr 47,000, identical to the viral product, that reacted with a specific monoclonal antibody.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cell Line
  • Cloning, Molecular
  • Coronaviridae / genetics*
  • DNA Restriction Enzymes
  • Escherichia coli / genetics
  • Genes*
  • Genes, Viral*
  • Molecular Sequence Data
  • RNA, Messenger / genetics
  • Saccharomyces cerevisiae / genetics*
  • Swine
  • Transcription, Genetic*
  • Transmissible gastroenteritis virus / genetics*
  • Transmissible gastroenteritis virus / isolation & purification

Substances

  • RNA, Messenger
  • DNA Restriction Enzymes