An improved method for the purification and refolding of r56-kDa proteins from Gilliam and Kato strains of Orientia tsutsugamushi

Ann N Y Acad Sci. 2003 Jun:990:375-85. doi: 10.1111/j.1749-6632.2003.tb07395.x.

Abstract

The immunodominant 56kDa outer membrane antigens from Orientia tsutsugamushi Kato and Gilliam strains were expressed as inclusion bodies (IBs) in E. coli. The IBs were purified and properly refolded with modifications of a previous procedure used for the production of Karp strain r56 antigen. A mixture of these three r56 proteins exhibited both high sensitivity and specificity for detection of O. tsutsugamushi antibodies by ELISA.

Publication types

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

MeSH terms

  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / isolation & purification*
  • Base Sequence
  • Chromatography, Gel
  • Chromatography, High Pressure Liquid
  • Cloning, Molecular
  • DNA Primers
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Molecular Weight
  • Orientia tsutsugamushi / classification
  • Orientia tsutsugamushi / genetics
  • Orientia tsutsugamushi / isolation & purification*
  • Protein Denaturation
  • Protein Folding
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism

Substances

  • Bacterial Proteins
  • DNA Primers
  • Recombinant Proteins