ArticleApplied and environmental microbiology2007
A rapid and efficient method for cloning genes of type II restriction-modification systems by use of a killer plasmid.
Article in Applied and environmental microbiology, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 8 citations in OpenAlex.
- Molecular Characterization of a Restriction Endonuclease PsaI fromInternational journal of molecular sciences · 2025Article
- Molecular basis for lethal cross-talk between two unrelated bacterial transcription factors - the regulatory protein of a restriction-modification system and the repressor of a defective prophage.Nucleic acids research · 2022Article
- Transcriptome analyses of cells carrying the Type II Csp231I restriction-modification system reveal cross-talk between two unrelated transcription factors: C protein and the Rac prophage repressor.Nucleic acids research · 2019Article
- Natural tuning of restriction endonuclease synthesis by cluster of rare arginine codons.Scientific reports · 2019Article
- Isospecific adenine DNA methyltransferases show distinct preferences towards DNA substrates.Scientific reports · 2018Article
- Plasmid stability analysis based on a new theoretical model employing stochastic simulations.PloS one · 2017Article
- Natural C-independent expression of restriction endonuclease in a C protein-associated restriction-modification system.Nucleic acids research · 2016Article
- Relaxed specificity of prokaryotic DNA methyltransferases results in DNA site-specific modification of RNA/DNA heteroduplexes.Journal of applied genetics · 2015Article
- Structural analysis of a novel class of R-M controller proteins: C.Csp231I fromJournal of molecular biology · 2011Article
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We present a method for cloning restriction-modification (R-M) systems that is based on the use of a lethal plasmid (pKILLER). The plasmid carries a functional gene for a restriction endonuclease having the same DNA specificity as the R-M system of interest. The first step is the standard preparation of a representative, plasmid-borne genomic library. Then this library is transformed with the killer plasmid. The only surviving bacteria are those which carry the gene specifying a protective DNA methyltransferase. Conceptually, this in vivo selection approach resembles earlier methods in which a plasmid library was selected in vitro by digestion with a suitable restriction endonuclease, but it is much more efficient than those methods. The new method was successfully used to clone two R-M systems, BstZ1II from Bacillus stearothermophilus 14P and Csp231I from Citrobacter sp. strain RFL231, both isospecific to the prototype HindIII R-M system.
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