Expression and purification of recombinant toxic-shock-syndrome toxin I

Jennifer L. Wahlsten, S. Ramakrishnan

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Toxic-shock-syndrome toxin I (TSSTI), an exotoxin produced by certain strains of Staphylococcus aureus, has been closely associated with the pathogenesis of toxic shock syndrome. Outside the context of its staphylococcal host, TSSTI may offer therapeutic uses. We report here a strategy for high-level expression and simplified purification of TSSTI. We have subcloned the coding region for TSSTI into a vector containing an inducible T7 promoter sequence and expressed the protein in an Escherichia coli host strain. The recombinant TSSTI protein contained ten sequential histidine residues (Histag) at its N-terminus, which enabled its efficient purification using nickel-agarose-affinity resin. Histag-TSSTI (H-TSSTI) was further purified to homogeneity using a size-exclusion column. By this system, 80 mg of highly purified H-TSSTI can be consistently obtained per litre of culture in under 3 days. H-TSSTI retained biological activity and was unaffected by the presence of the Histag, as measured in lymphocyte proliferation assays.

Original languageEnglish (US)
Pages (from-to)155-159
Number of pages5
JournalBiotechnology and Applied Biochemistry
Volume24
Issue number2
StatePublished - Oct 14 1996
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Molecular Medicine
  • Biomedical Engineering
  • Applied Microbiology and Biotechnology
  • Drug Discovery
  • Process Chemistry and Technology

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