title: Curing a large endogenous plasmid by single substitution of a partitioning gene.
abstract: To investigate whether plasmid-free cells of pathogenic Escherichia coli can be isolated by disrupting a single gene in an endogenous plasmid without further treatment, the effect of the disruption of partitioning genes on the inheritance of the endogenous plasmid pUTI89 of the uropathogenic E. coli strain UTI89 was studied. We found that mutation of parB, which encodes a type Ib partitioning protein, could cause loss of the endogenous plasmid at a ratio of about 1%. Clones derived from parB mutants, identified by antibiotic sensitivity, were all plasmid free. Plasmid instability caused by the parB mutation was found to correlate with a negative effect on host cell growth. Thus, in this pathogenic E. coli, an endogenous plasmid as large as 114 kbp could be cured effectively by targeting a single type Ib partitioning gene followed by passaging, which may facilitate further investigations on the function of endogenous plasmids in their natural hosts.
authors.affiliation: State Key Lab of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001 Harbin, China; Microbiology Research Group, Faculty of Science and Bio-engineering Sciences, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium.
authors.forename: Ningning
authors.initials: N
authors.lastname: Song
authors.affiliation: State Key Laboratory for Oncogenes and Related Genes, Division of Gastroenterology and Hepatology, Renji Hospital, 200127 Shanghai, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Xu
authors.affiliation: State Key Lab of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 150001 Harbin, China; Viral Genetics Research Group, Faculty of Science and Bio-engineering Sciences, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium. Electronic address: zhaoli.li80@qq.com.
authors.forename: Zhaoli
authors.initials: Z
authors.lastname: Li
authors.affiliation: Viral Genetics Research Group, Faculty of Science and Bio-engineering Sciences, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Hernalsteens
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D019915:DNA Primase / Q000235:genetics*; D004251:DNA Transposable Elements / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D029968:Escherichia coli Proteins / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics; D007612:Kanamycin / Q000494:pharmacology; D009154:Mutation / Q000235:genetics; D010957:Plasmids / Q000235:genetics*
chemical_list: D000900:Anti-Bacterial Agents; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D007612:Kanamycin; D019915:DNA Primase; C121791:dnaG protein, E coli
keywords: Escherichia coli; Lambda recombination system; Partitioning gene; Plasmid curing; Plasmid-free cells; Transposon insertion mutants