Production of recombinant porin from Y. pseudotuberculosis in a water-soluble form for pseudotuberculosis diagnostics

Biol Chem. 2017 Oct 26;398(11):1229-1236. doi: 10.1515/hsz-2017-0142.

Abstract

OmpF porin from the outer membrane of Yersinia pseudotuberculosis was cloned into pET-40b(+) plasmid. Using E. coli Rosetta (DE3) strain, MX medium, IPTG concentration of 0.2 mm and post-induction cultivation at 14°C overnight allowed us to obtain a water-soluble form of the recombinant protein (rs-OmpF). Rs-OmpF was shown to have the ordered spatial structure at the levels of secondary and tertiary structure. Rs-OmpF was found to be effective as diagnostic antigen in ELISA for pseudotuberculosis diagnostics.

Keywords: E. coli; ELISA; membrane proteins; porin; soluble recombinant proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Porins / biosynthesis*
  • Porins / genetics
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Solubility
  • Water / chemistry*
  • Yersinia pseudotuberculosis / chemistry*
  • Yersinia pseudotuberculosis / genetics
  • Yersinia pseudotuberculosis Infections / diagnosis*
  • Yersinia pseudotuberculosis Infections / microbiology

Substances

  • OmpF protein
  • Porins
  • Recombinant Proteins
  • Water