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

Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review

Autoren

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

Externe Organisationen

  • Helmholtz-Zentrum für Infektionsforschung GmbH (HZI)
  • Universidad Autónoma de Barcelona (UAB)
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Details

OriginalspracheEnglisch
Seiten (von - bis)327-334
Seitenumfang8
FachzeitschriftFEMS microbiology letters
Jahrgang177
Ausgabenummer2
PublikationsstatusVeröffentlicht - 15 Aug. 1999
Extern publiziertJa

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.

ASJC Scopus Sachgebiete

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Heat-inactivation of plasmid-encoded CI857 repressor induces gene expression from Ind - lambda prophage in recombinant Escherichia coli. / Hoffmann, Frank; Arís, Anna; Carbonell, Xavier et al.
in: FEMS microbiology letters, Jahrgang 177, Nr. 2, 15.08.1999, S. 327-334.

Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review

Hoffmann F, Arís A, Carbonell X, Rohde M, Corchero JL, Rinas U et al. Heat-inactivation of plasmid-encoded CI857 repressor induces gene expression from Ind - lambda prophage in recombinant Escherichia coli. FEMS microbiology letters. 1999 Aug 15;177(2):327-334. doi: 10.1016/S0378-1097(99)00334-1
Hoffmann, Frank ; Arís, Anna ; Carbonell, Xavier et al. / Heat-inactivation of plasmid-encoded CI857 repressor induces gene expression from Ind - lambda prophage in recombinant Escherichia coli. in: FEMS microbiology letters. 1999 ; Jahrgang 177, Nr. 2. S. 327-334.
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title = "Heat-inactivation of plasmid-encoded CI857 repressor induces gene expression from Ind - lambda prophage in recombinant Escherichia coli",
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.",
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author = "Frank Hoffmann and Anna Ar{\'i}s and Xavier Carbonell and Manfred Rohde and Corchero, {Jos{\'e} L.} and Ursula Rinas and Antonio Villaverde",
note = "Funding Information: We are indebted to M. Defais for generously providing strain GE864 and FL8641/pFL352 and to M. Blanco for strain GC4581 and helpful discussions. An {\textquoteleft}Acciones Integradas{\textquoteright} Spanish-German co-operation programme was supporting part of the work: HA96-0028, HA97-0136 (MEC) and 314-A1-e-dr (DAAD). Work in Barcelona was also supported by CICYT (Grant BIO98-0527) and by the Fundaci{\'o} Maria Francesca de Roviralta, Spain. A.A. and J.L.C. were recipients of predoctoral fellowships from the Ministerio de Educaci{\'o}n y Cultura, Spain.",
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Download

TY - JOUR

T1 - Heat-inactivation of plasmid-encoded CI857 repressor induces gene expression from Ind - lambda prophage in recombinant Escherichia coli

AU - Hoffmann, Frank

AU - Arís, Anna

AU - Carbonell, Xavier

AU - Rohde, Manfred

AU - Corchero, José L.

AU - Rinas, Ursula

AU - Villaverde, Antonio

N1 - Funding Information: We are indebted to M. Defais for generously providing strain GE864 and FL8641/pFL352 and to M. Blanco for strain GC4581 and helpful discussions. An ‘Acciones Integradas’ Spanish-German co-operation programme was supporting part of the work: HA96-0028, HA97-0136 (MEC) and 314-A1-e-dr (DAAD). Work in Barcelona was also supported by CICYT (Grant BIO98-0527) and by the Fundació Maria Francesca de Roviralta, Spain. A.A. and J.L.C. were recipients of predoctoral fellowships from the Ministerio de Educación y Cultura, Spain.

PY - 1999/8/15

Y1 - 1999/8/15

N2 - 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.

AB - 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.

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KW - CI857 repressor

KW - Lambda bacteriophage

KW - Recombinant protein

KW - Structural gene

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