Peptides from the SARS-associated coronavirus as tags for protein expression and purification

Protein Expr Purif. 2008 Oct;61(2):138-41. doi: 10.1016/j.pep.2008.04.011. Epub 2008 May 9.

Abstract

Protein tagging with a peptide is a commonly used technique to facilitate protein detection and to carry out protein purification. Flexibility with respect to the peptide tag is essential since no single tag suites all purposes. This report describes the usage of two short peptides from the SARS-associated coronavirus nucleocapsid (SARS-N) protein as protein tags. Plasmids for the generation of tagged proteins were generated by ligating synthetic oligonucleotides for the peptide-coding regions downstream of the protein coding sequence. The data show recognition of prokaryotically expressed HIV-1 Gag/p24 fusion protein by Western blot and efficient affinity purification using monoclonal antibodies against the tags. The SARS peptide antibody system described presents an alternative tagging opportunity in the growing field of protein science.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / chemistry
  • Blotting, Western
  • Chromatography, Affinity / methods
  • Coronavirus Nucleocapsid Proteins
  • Electrophoresis, Polyacrylamide Gel / methods
  • Escherichia coli / chemistry
  • Escherichia coli / metabolism
  • Nucleocapsid Proteins* / biosynthesis
  • Nucleocapsid Proteins* / chemistry
  • Nucleocapsid Proteins* / isolation & purification
  • Peptide Mapping
  • Peptides / chemistry*
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / isolation & purification
  • Severe acute respiratory syndrome-related coronavirus / chemistry*

Substances

  • Antibodies, Monoclonal
  • Coronavirus Nucleocapsid Proteins
  • Nucleocapsid Proteins
  • Peptides
  • Recombinant Fusion Proteins