title: [Construction of recombinant plasmid with Porphyromonas gingivalis FimA deficiency].
abstract: OBJECTIVE
  To construct the recombinant plasmid pPHU281_A_Spec_B, which knock out Porphyrmonas gingivalis (Pg) FimA gene.
  
  
  METHODS
  Genomic DNA was extracted from PgATCC33277 which was cultured in anaerobic condition. The upstream and downstream gene of FimA was cloned from Pg genenomic DNA with specific restriction sites by polymerase chain reaction. Suicide vector pPHU281 was inserted by three fragments: upstream, downstream of FimA gene and spectinomycin resistance gene. The recombinant plasmid was confirmed by electrophoresis and sequenced after amplification in compentent cells DH-5α.
  
  
  RESULTS
  The gene sequence was identified by DNA sequencing analysis. The recombinant plasmid pPHU281_A_Spec_B was successfully constructed.
  
  
  CONCLUSIONS
  The recombinant plasmid pPHU281_A_Spec_B was constructed, which may be used for the constructon of FimA deficient Pg.
authors.affiliation: Department of Periodontology, School of Stomatology, Nanjing University & Nanjing Stomatological Hospital, Nanjing, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Yang
authors.forename: Kuan-Yu
authors.initials: KY
authors.lastname: Li
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
authors.forename: Juan
authors.initials: J
authors.lastname: Wu
authors.forename: Wei-Bin
authors.initials: WB
authors.lastname: Sun
mesh_terms: D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D036781:Fimbriae Proteins / Q000235:genetics*; D055786:Gene Knockout Techniques; D005798:Genes, Bacterial; D005822:Genetic Vectors; D010957:Plasmids / Q000235:genetics*; D016966:Porphyromonas gingivalis / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: D004269:DNA, Bacterial; D011994:Recombinant Proteins; C063055:fimbrillin; D036781:Fimbriae Proteins