title: Identification of Cj1051c as a major determinant for the restriction barrier of Campylobacter jejuni strain NCTC11168.
abstract: Campylobacter jejuni is a leading cause of human diarrheal illness in the world, and research on it has benefitted greatly by the completion of several genome sequences and the development of molecular biology tools. However, many hurdles remain for a full understanding of this unique bacterial pathogen. One of the most commonly used strains for genetic work with C. jejuni is NCTC11168. While this strain is readily transformable with DNA for genomic recombination, transformation with plasmids is problematic. In this study, we have identified a determinant of this to be cj1051c, predicted to encode a restriction-modification type IIG enzyme. Knockout mutagenesis of this gene resulted in a strain with a 1,000-fold-enhanced transformation efficiency with a plasmid purified from a C. jejuni host. Additionally, this mutation conferred the ability to be transformed by plasmids isolated from an Escherichia coli host. Sequence analysis suggested a high level of variability of the specificity domain between strains and that this gene may be subject to phase variation. We provide evidence that cj1051c is active in NCTC11168 and behaves as expected for a type IIG enzyme. The identification of this determinant provides a greater understanding of the molecular biology of C. jejuni as well as a tool for plasmid work with strain NCTC11168.
authors.affiliation: Department of Biology, Drexel University, Philadelphia, Pennsylvania, USA.
authors.forename: Jeffrey P
authors.initials: JP
authors.lastname: Holt
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Grant
authors.forename: Christopher
authors.initials: C
authors.lastname: Coward
authors.forename: Duncan J
authors.initials: DJ
authors.lastname: Maskell
authors.forename: Jennifer J
authors.initials: JJ
authors.lastname: Quinlan
mesh_terms: D016123:Campylobacter jejuni / Q000235:genetics*; D004262:DNA Restriction Enzymes / Q000235:genetics* / Q000378:metabolism*; D015280:DNA Restriction-Modification Enzymes / Q000235:genetics*; D004269:DNA, Bacterial / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D055786:Gene Knockout Techniques; D006868:Hydrolysis; D010957:Plasmids; D014169:Transformation, Bacterial
chemical_list: D015280:DNA Restriction-Modification Enzymes; D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes