Cloning and polymorphism analysis of prtH gene from Porphyromonas gingivalis.
- Author:
Ying ZHENG
1
;
Sheng-hui YANG
;
Wei ZHOU
;
Chun-mei ZHANG
;
Fu-ping ZHANG
;
Xiao-ping DONG
Author Information
- Publication Type:Journal Article
- MeSH: Bacterial Proteins; Blotting, Southern; Cloning, Molecular; Cysteine Endopeptidases; genetics; DNA, Bacterial; genetics; metabolism; Deoxyribonuclease BamHI; metabolism; Deoxyribonuclease HindIII; metabolism; Polymorphism, Genetic; Porphyromonas gingivalis; genetics; Species Specificity
- From: Chinese Journal of Stomatology 2003;38(1):27-30
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSUsing 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.
RESULTSRecombinant 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.
CONCLUSIONSDifferent 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.
