Uses of β-galactosidase tag in on-line monitoring production of fusion proteins and gene expression in Escherichia coli

A. Benito, F. Valero, J. Lafuente, M. Vidal, J. Cairo, C. Solà, A. Villaverde

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21 Citations (Scopus)

Abstract

A simple method for monitoring and quantifying automatically the production by fermentation of β-galactosidase fusion proteins, making use of the remaining activity of the β-galactosidase part, is considered. A hybrid protein carrying the major antigenic domain of foot-and-mouth disease virus C1 joined at the N-terminus of β-galactosidase has been expressed in Escherichia coli. The yield of the chimeric protein has been monitored by flow injection analysis (FIA) during batch fermentations at laboratory scale, and a high correlation between values of product concentration from FIA and from immunological quantizations has been obtained. Because of the possibility of employing FIA in large-scale experiments, and the high sampling frequency, versatility, and reproducibility offered by this method, we propose FIA as a general, simple, quick, flexible, and reliable instrument for both monitoring the yield of recombinant proteins produced industrially, and performing basic research at laboratory scale. © 1993.
Original languageEnglish
Pages (from-to)66-71
JournalEnzyme and Microbial Technology
Volume15
DOIs
Publication statusPublished - 1 Jan 1993

Keywords

  • FMDV
  • Flow Injection Analysis (FIA)
  • fermentation monitoring
  • gene expression
  • recombinant proteins
  • β-galactosidase

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