[Cloning and polymorphism analysis of prtH gene from Porphyromonas gingivalis].
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.
Zheng
Ying
Y
Beijing Institute of Dental Research, Beijing Hospital for Stomatology, Capital University of Medical Sciences, Beijing 100050, China.
Yang
Sheng-hui
SH
Zhou
Wei
W
Zhang
Chun-mei
CM
Zhang
Fu-ping
FP
Dong
Xiao-ping
XP
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
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D015245:Deoxyribonuclease BamHI; D015247:Deoxyribonuclease HindIII; D003546:Cysteine Endopeptidases; C089744:PrtH protein, bacteria