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

Claudia Saavedra, Enrique González, Claudio Vásquez

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)391-397
Number of pages7
JournalBiochemistry and Molecular Biology International
Volume44
Issue number2
Publication statusPublished - Feb 1998

Fingerprint

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

Keywords

  • B. stearothermophilusV
  • Isoschizomer
  • Restriction-modification system
  • Thermostable

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

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

In: Biochemistry and Molecular Biology International, Vol. 44, No. 2, 02.1998, p. 391-397.

Research output: Contribution to journalArticle

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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.

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