title: [Construction PG0839 gene-defective mutant of Porphyromonas gingivalis].
abstract: OBJECTIVE
  In order to determine the function of PG0839 gene from Porphyromonas gingivalis (P. gingivalis) W83 strains, we intended to create a mutant in the PG0839 gene by homologous recombination.
  
  
  METHODS
  1 584bp PG0839 gene fragment was amplified, digested by BamH I and EcoR I, purified and ligated to pUC19. The recombinant plasmid was designated as pPG0839-1. The erm cassette (2 101 bp) was inserted into the EcoR V restriction site of the PG0839 gene. The resultant recombinant plasmid, pPG0839-2, was used as a donor in the electroporation of P. gingivalis W83. After electroporated and selected on erythromycin brain heart infusion plates, a single colony was collected and designated as PG0839 gene-defective mutant.
  
  
  RESULTS
  A mutant in PG0839 gene was created by insertional inactivation, and inactivation of PG0839 gene was confirmed by restriction endonuclease digestive, sequencing, polymerase chain reaction (PCR) and reverse transcription PCR.
  
  
  CONCLUSIONS
  A PG0839 gene-defective mutant was created successfully.
authors.affiliation: Dept. of Periodontology, School of Stomatology, China Medical University, Shenyang 110002, China.
authors.forename: Jingbo
authors.initials: J
authors.lastname: Liu
authors.forename: Yaping
authors.initials: Y
authors.lastname: Pan
authors.forename: Chen
authors.initials: C
authors.lastname: Li
authors.forename: Li
authors.initials: L
authors.lastname: Lin
authors.forename: Ming
authors.initials: M
authors.lastname: Zhong
mesh_terms: D001483:Base Sequence; D005798:Genes, Bacterial; D016133:Polymerase Chain Reaction; D016966:Porphyromonas gingivalis