title: Insights into the genetic organization of the Corynebacterium diphtheriae erythromycin resistance plasmid pNG2 deduced from its complete nucleotide sequence.
abstract: The complete nucleotide sequence of the erythromycin resistance plasmid pNG2 from the human pathogen Corynebacterium diphtheriae S601 was determined. The plasmid has a total size of 15,100 bp and contains at least 17 coding regions. Comparative genomics identified conserved motifs within replication initiator proteins of corynebacterial plasmids and a novel nucleotide sequence feature, termed 22-bp box, located downstream of the repA gene. The erythromycin resistance determinant erm(X) is flanked by inverted repeats of the novel insertion sequence IS3504, which may be responsible for a spontaneous deletion of the antibiotic resistance gene region. Furthermore, pNG2 encodes a putative conjugative relaxase, a membrane protein of the natural resistance-associated macrophage protein (Nramp) family and a protein with Nudix hydrolase signature. Expression of the predicted coding regions of pNG2 in Escherichia coli JM109 was demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) assays. The detailed annotation of the entire pNG2 sequence provided genetic information regarding its molecular evolution and its role in dissemination of antibiotic resistance genes by horizontal gene transfer.
authors.affiliation: Zentrum für Genomforschung, Universität Bielefeld, Universitätsstrasse 25, D-33615 Bielefeld, Germany. andreas.tauch@genetik.uni-bielfeld.de
authors.forename: Andreas
authors.initials: A
authors.lastname: Tauch
authors.forename: Nicole
authors.initials: N
authors.lastname: Bischoff
authors.forename: Iris
authors.initials: I
authors.lastname: Brune
authors.forename: Jörn
authors.initials: J
authors.lastname: Kalinowski
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D017124:Conserved Sequence; D003353:Corynebacterium diphtheriae / Q000187:drug effects / Q000235:genetics*; D004265:DNA Helicases / Q000235:genetics; D004254:DNA Nucleotidyltransferases / Q000235:genetics; D004251:DNA Transposable Elements; D021901:DNA, Intergenic; D004268:DNA-Binding Proteins; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004917:Erythromycin / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics; D023061:Gene Order; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*; D011506:Proteins / Q000235:genetics; D011755:Pyrophosphatases / Q000235:genetics; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D015534:Trans-Activators; D000096964:Nudix Hydrolases
chemical_list: D001426:Bacterial Proteins; D004265:DNA Helicases; D004254:DNA Nucleotidyltransferases; D004251:DNA Transposable Elements; D021901:DNA, Intergenic; D004268:DNA-Binding Proteins; D004917:Erythromycin; D011506:Proteins; D011755:Pyrophosphatases; D015534:Trans-Activators; C045264:replication initiator protein; C110340:DNA relaxase; C422942:nudix hydrolase Orf135; D000096964:Nudix Hydrolases