title: [Cloning and polymorphism analysis of prtH gene from Porphyromonas gingivalis].
abstract: OBJECTIVE
  To clone the prtH gene from Porphyromonas gingivalis (P.g) ATCC 33277 and analyze the polymorphism of prtH gene from 5 strains of P.g in order to explore the relationship between P.g and periodontitis.
  
  
  METHODS
  Using PCR, the prtH was amplified and cloned into pGEM-T vector. To illustrate the prtH polymorphism among P.g strains, the genomic DNAs were extracted and screened by PCR with three pairs of specific primers, dot blot and Southern blot hybridization using the biotin-labeled prtH sequence as probe.
  
  
  RESULTS
  Recombinant DNA pGEM-T- prtH was verified by restriction endonuclease and sequence assay. Strain W 381 and ATCC 33277 showed the identical results in PCR and hybridization assays, whereas strain ATCC 49417 and 14-3-2 revealed individual hybridization patterns. Strain 47A-1 seemed even not to contain prtH gene.
  
  
  CONCLUSIONS
  Different prtH gene sequences exist in different P.g strains. This polymorphism may indicate various potential virulent effects during the infection and pathogenesis. Established PCR protocol is sensitive for identification of prtH gene.
authors.affiliation: Beijing Institute of Dental Research, Beijing Hospital for Stomatology, Capital University of Medical Sciences, Beijing 100050, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zheng
authors.forename: Sheng-hui
authors.initials: SH
authors.lastname: Yang
authors.forename: Wei
authors.initials: W
authors.lastname: Zhou
authors.forename: Chun-mei
authors.initials: CM
authors.lastname: Zhang
authors.forename: Fu-ping
authors.initials: FP
authors.lastname: Zhang
authors.forename: Xiao-ping
authors.initials: XP
authors.lastname: Dong
mesh_terms: D001426:Bacterial Proteins; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D015245:Deoxyribonuclease BamHI / Q000378:metabolism; D015247:Deoxyribonuclease HindIII / Q000378:metabolism; D011110:Polymorphism, Genetic; D016966:Porphyromonas gingivalis / Q000235:genetics*; D013045:Species Specificity
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D015245:Deoxyribonuclease BamHI; D015247:Deoxyribonuclease HindIII; D003546:Cysteine Endopeptidases; C089744:PrtH protein, bacteria