Heat-inactivation of plasmid-encoded CI857 repressor induces gene expression from Ind - lambda prophage in recombinant Escherichia coli

Frank Hoffmann, Anna Arís, Xavier Carbonell, Manfred Rohde, José L. Corchero, Ursula Rinas, Antonio Villaverde

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4 Citations (Scopus)

Abstract

We have observed significant cell lysis upon temperature up-shift of recombinant Escherichia coli cultures harboring CI857-repressed lambda-based expression vectors. This event, that becomes evident about 30-40 min after the heat shock, takes place when using the lambda promoter system in Ind - lysogenic strains, but not in others commonly employed for recombinant gene expression. These results strongly suggest that the thermosensitive CI857 repressor, encoded by the expression vector, competes with CI Ind - molecules for binding to the prophage operator region, allowing for expression of lytic genes from the integrated Ind - viral genome upon temperature up-shift. Transcription of viral lytic genes does not include unspecific expression of a reporter sulA::lacZ gene fusion carried in the prophage genome. These results prompt, however, to carefully evaluate the limitations of expression systems based on p(L)/p(R)-CI857 in bacterial strains modified through lambda Ind - gene transfer vehicles. Copyright (C) 1999 Federation of European Microbiological Societies.
Original languageEnglish
Pages (from-to)327-334
JournalFEMS Microbiology Letters
Volume177
DOIs
Publication statusPublished - 15 Aug 1999

Keywords

  • CI857 repressor
  • Cell lysis
  • Lambda bacteriophage
  • Recombinant protein
  • Structural gene

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