Details
| Original language | English |
|---|---|
| Pages (from-to) | 327-334 |
| Number of pages | 8 |
| Journal | FEMS microbiology letters |
| Volume | 177 |
| Issue number | 2 |
| Publication status | Published - 15 Aug 1999 |
| Externally published | Yes |
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.
Keywords
- Cell lysis, CI857 repressor, Lambda bacteriophage, Recombinant protein, Structural gene
ASJC Scopus subject areas
- Immunology and Microbiology(all)
- Microbiology
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
- Biochemistry, Genetics and Molecular Biology(all)
- Genetics
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In: FEMS microbiology letters, Vol. 177, No. 2, 15.08.1999, p. 327-334.
Research output: Contribution to journal › Article › Research › peer review
}
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.
KW - Cell lysis
KW - CI857 repressor
KW - Lambda bacteriophage
KW - Recombinant protein
KW - Structural gene
UR - http://www.scopus.com/inward/record.url?scp=0345161776&partnerID=8YFLogxK
U2 - 10.1016/S0378-1097(99)00334-1
DO - 10.1016/S0378-1097(99)00334-1
M3 - Article
C2 - 10474200
AN - SCOPUS:0345161776
VL - 177
SP - 327
EP - 334
JO - FEMS microbiology letters
JF - FEMS microbiology letters
SN - 0378-1097
IS - 2
ER -