Expression of reovirus type 3 (Dearing) sigma 1 and sigma s polypeptides in Escherichia coli

J Gen Virol. 1987 Jan:68 ( Pt 1):135-45. doi: 10.1099/0022-1317-68-1-135.

Abstract

The reovirus S1 gene codes for two polypeptides: sigma 1 and sigma s. In order to characterize the structure and function of the sigma 1 polypeptide, we have expressed the sigma 1 protein in Escherichia coli. The S1 gene from mammalian reovirus type 3 (Dearing strain) and the variant K strain were subcloned into an expression vector containing the tac (trp-lac) promoter designed to express foreign gene products in E. coli efficiently. The hybrid plasmids, upon induction with isopropyl-beta-D-thiogalactopyranoside, expressed two polypeptides that were detected by [35S]methionine labelling. One of the induced proteins had a relative molecular mass (Mr) of approx. 46,000 and corresponded to sigma 1, as shown by immunoprecipitation with goat anti-reovirus antibody and a monoclonal antibody against sigma 1. The second induced protein had a Mr of approx. 12,000 and was very similar to sigma s as judged by comparative tryptic peptide map analysis. Protein sigma 1 produced in E. coli was shown to be functional as judged from its ability to bind to mouse L fibroblasts.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Capsid Proteins*
  • Cloning, Molecular
  • Escherichia coli / genetics*
  • Genes*
  • Genes, Viral*
  • Genetic Variation
  • Genetic Vectors
  • L Cells / metabolism
  • Mammalian orthoreovirus 3 / genetics*
  • Mice
  • Plasmids
  • Reoviridae / genetics*
  • Transcription, Genetic
  • Viral Proteins / genetics*

Substances

  • Capsid Proteins
  • Viral Proteins
  • sigma 1 protein, reovirus