A Rapid and Efficient Method for Cloning Genes of Type II Restriction-modification Systems by Use of a Killer Plasmid

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 2007 May 1

PMID 17468281

Citations 8

Authors

Iwona Mruk

Tadeusz Kaczorowski

Affiliations

Soon will be listed here.

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.

Citing Articles

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.

Wisniewska A, Wons E, Potrykus K, Hinrichs R, Gucwa K, Graumann P Nucleic Acids Res. 2022; 50(19):10964-10980.

PMID: 36271797 PMC: 9638921. DOI: 10.1093/nar/gkac914.

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.

Negri A, Jakalski M, Szczuka A, Pryszcz L, Mruk I Nucleic Acids Res. 2019; 47(18):9542-9556.

PMID: 31372643 PMC: 6765115. DOI: 10.1093/nar/gkz665.

Natural tuning of restriction endonuclease synthesis by cluster of rare arginine codons.

Mruk I, Kaczorowski T, Witczak A Sci Rep. 2019; 9(1):5808.

PMID: 30967604 PMC: 6456624. DOI: 10.1038/s41598-019-42311-w.

Isospecific adenine DNA methyltransferases show distinct preferences towards DNA substrates.

Wons E, Mruk I, Kaczorowski T Sci Rep. 2018; 8(1):8243.

PMID: 29844340 PMC: 5974420. DOI: 10.1038/s41598-018-26434-0.

Plasmid stability analysis based on a new theoretical model employing stochastic simulations.

Werbowy O, Werbowy S, Kaczorowski T PLoS One. 2017; 12(8):e0183512.

PMID: 28846713 PMC: 5573283. DOI: 10.1371/journal.pone.0183512.

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