Abstract
The geneshsdM andhsdS for M.EcoKI modification methyltrasferase and the complete set ofhsdR,hsdM andhsdS genes coding for R.EcoKI restriction endonuclease, both with and without a temperature-sensitive (ts) mutation inhsdS gene, were cloned in pBR322 plasmid and introduced intoE. coli C (a strain without a natural restriction-modification (R-M) system). The strains producing only the methyltransferase, or together with the endonuclease, were thus obtained. ThehsdS ts-1 mutation, mapped previously in the distal variable region of thehsdS gene with C1 245-T transition has no effect on the R-M phenotype expressed from cloned genes in bacteria grown at 42°C. In clones transformed with the wholehsd region an alleviation of R-M functions was observed immediately after the transformation, but after subculture the transformants expressed the wild-type R-M phenotype irrespective of whether the wild-type or the mutanthsdS allele was present in the hybrid plasmid. Simultaneous overproduction of HsdS and HsdM subunits impairs the ts effect of thehsdS ts-1 mutation on restriction and modification.
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Abbreviations
- Amp:
-
ampicillin
- EtdBr:
-
ethidium bromide
- ts:
-
temperature-sensitive
- EOP:
-
efficiency of plating
- R-M:
-
restriction and modification
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Weiserová, M., Janščák, P., Zinkevich, V. et al. Overproduction of the hsd subunits leads to the loss of temperature-sensitive restriction and modification phenotype. Folia Microbiol 39, 452–458 (1994). https://doi.org/10.1007/BF02814061
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DOI: https://doi.org/10.1007/BF02814061