Immunological characterization of the sheep prion protein expressed as fusion proteins in Escherichia coli

FEMS Immunol Med Microbiol. 1999 Sep;25(4):379-84. doi: 10.1111/j.1574-695X.1999.tb01363.x.

Abstract

The prion protein (PrP) from sheep was produced in large quantities of entire protein in Escherichia coli after fusion with a carboxy-terminal hexahistidine sequence. In contrast, amino-terminal fusion with glutathione S-transferase (GST) revealed a high susceptibility toward cleavage of the protein. Both recombinant proteins were recognised, at variable levels, in Western blots using a panel of antibodies against the 40-56, 89-104, 98-113 and 112-115 sequences of the prion protein, similarly to the abnormal prion protein extracted from scrapie-infected sheep. Interestingly, monoclonal antibody 3F4 was found to react with these three proteins in Western blot.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cloning, Molecular
  • Escherichia coli
  • Gene Expression
  • Glutathione Transferase / genetics
  • Glutathione Transferase / metabolism
  • Histidine
  • Prions / genetics
  • Prions / immunology*
  • Prions / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Recombinant Fusion Proteins / metabolism
  • Sheep

Substances

  • Prions
  • Recombinant Fusion Proteins
  • Histidine
  • Glutathione Transferase