Polyoma virus major capsid protein, VP1. Purification after high level expression in Escherichia coli

J Biol Chem. 1985 Oct 15;260(23):12803-9.

Abstract

We have expression-cloned in Escherichia coli the major polyoma virus capsid protein, VP1. Under the inducible control of the hybrid tac promoter, VP1 constituted between 2 and 3% of the total host cell protein. The expressed VP1 was purified to near homogeneity with initial yields to 10%. Optimal expression was temperature-dependent, and significant intracellular degradation could be demonstrated. The final product was obtained as one predominant isoelectric focusing species, without the pattern of post-translational modification seen in virus-infected eukaryotic cells. The purified VP1 from E. coli will be useful as a substrate for the purification of VP1 modification enzymes and in the study of inter-VP1 oligomerization.

Publication types

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

MeSH terms

  • Cloning, Molecular
  • DNA Restriction Enzymes
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics*
  • Fractional Precipitation
  • Immunosorbent Techniques
  • Isoelectric Focusing
  • Molecular Weight
  • Plasmids
  • Polyomavirus / genetics*
  • Promoter Regions, Genetic
  • Transcription, Genetic
  • Viral Proteins / genetics*
  • Viral Proteins / isolation & purification
  • Viral Structural Proteins

Substances

  • Viral Proteins
  • Viral Structural Proteins
  • DNA Restriction Enzymes