Studies on the heterologous expression of BstVI restriction endonuclease in Escherichia coli

Claudia Saavedra, Enrique González, Claudio Vásquez

Resultado de la investigación: Article

3 Citas (Scopus)

Resumen

Bacterial restriction and modification systems must be regulated to avoid self-restriction. It is generally accepted that cognate DNA methyltransferases normally protects both, the host's chromosome and extrachromosomal elements from the activity of their endonuclease counterparts. When the bstVIRM genes from Bacillus stearothermophilusV were subcloned into Escherichia coli, several clones exhibiting a r+m- phenotype were originated. The present work was undertaken to analyze the possibility that mechanisms other than DNA methylation could account for the viability of these cells. No evidence was found for an inhibitory agent or endonuclease compartmentation. In vivo experiments showed that h phage multiplication was poorly restricted by the heterologous enzyme. The restricting activity against the incoming phage increased however when phage adsortion was performed at higher temperatures. Analogous experiments in which a DNA-repair deficient strain was used as a host for the thermophilic R-M system suggested, to some extent, the participation of the repair machinery in the viability of r+m- clones.

Idioma originalEnglish
Páginas (desde-hasta)391-397
Número de páginas7
PublicaciónBiochemistry and Molecular Biology International
Volumen44
N.º2
EstadoPublished - feb 1998

Huella dactilar

Bacteriophages
Escherichia coli
Endonucleases
Repair
Clone Cells
DNA Restriction-Modification Enzymes
DNA Repair-Deficiency Disorders
DNA
Methyltransferases
Bacilli
DNA Methylation
Chromosomes
Bacillus
Machinery
Cell Survival
Genes
Experiments
Cells
Phenotype
Temperature

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Genetics

Citar esto

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Studies on the heterologous expression of BstVI restriction endonuclease in Escherichia coli. / Saavedra, Claudia; González, Enrique; Vásquez, Claudio.

En: Biochemistry and Molecular Biology International, Vol. 44, N.º 2, 02.1998, p. 391-397.

Resultado de la investigación: Article

TY - JOUR

T1 - Studies on the heterologous expression of BstVI restriction endonuclease in Escherichia coli

AU - Saavedra, Claudia

AU - González, Enrique

AU - Vásquez, Claudio

PY - 1998/2

Y1 - 1998/2

N2 - Bacterial restriction and modification systems must be regulated to avoid self-restriction. It is generally accepted that cognate DNA methyltransferases normally protects both, the host's chromosome and extrachromosomal elements from the activity of their endonuclease counterparts. When the bstVIRM genes from Bacillus stearothermophilusV were subcloned into Escherichia coli, several clones exhibiting a r+m- phenotype were originated. The present work was undertaken to analyze the possibility that mechanisms other than DNA methylation could account for the viability of these cells. No evidence was found for an inhibitory agent or endonuclease compartmentation. In vivo experiments showed that h phage multiplication was poorly restricted by the heterologous enzyme. The restricting activity against the incoming phage increased however when phage adsortion was performed at higher temperatures. Analogous experiments in which a DNA-repair deficient strain was used as a host for the thermophilic R-M system suggested, to some extent, the participation of the repair machinery in the viability of r+m- clones.

AB - Bacterial restriction and modification systems must be regulated to avoid self-restriction. It is generally accepted that cognate DNA methyltransferases normally protects both, the host's chromosome and extrachromosomal elements from the activity of their endonuclease counterparts. When the bstVIRM genes from Bacillus stearothermophilusV were subcloned into Escherichia coli, several clones exhibiting a r+m- phenotype were originated. The present work was undertaken to analyze the possibility that mechanisms other than DNA methylation could account for the viability of these cells. No evidence was found for an inhibitory agent or endonuclease compartmentation. In vivo experiments showed that h phage multiplication was poorly restricted by the heterologous enzyme. The restricting activity against the incoming phage increased however when phage adsortion was performed at higher temperatures. Analogous experiments in which a DNA-repair deficient strain was used as a host for the thermophilic R-M system suggested, to some extent, the participation of the repair machinery in the viability of r+m- clones.

KW - B. stearothermophilusV

KW - Isoschizomer

KW - Restriction-modification system

KW - Thermostable

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