Heterogeneity of Porphyromonas gingivalis strains on fimbrillin gene locus by restriction fragment length polymorphism analysis.

Y J Zhang
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Abstract

The adherence of Porphyromonas gingivalis (P. gingivalis) to the periodontal tissue may be an initial step in the pathogenesis of periodontal disease, and fimbriae is believed to play an important role in such processes. However, the heterogeneity of the size, sequence and antigenic reactivity of the fimbriae have been reported. The aim of the present study was to investigate the heterogeneity of P. gingivalis strains on the fimbrillin gene locus by RFLPs analysis. Fourty-seven P. gingivalis strains including 5 reference strains were used in this study. The plasmid (pUC13Bg 12.1) with the insert of fimA381 was modified with bisulfite and used as the probe. The genomic DNAs from P. gingivalis were digested with the restriction endonuclease SacI and PstI, electrophoresed, transferred and hybridized with the DNA probe. All Sac I digests generated one major band and the band size was almost the same (ca 2.5-kb), except strain W50. The Pst I digests showed one or two major bands and could be divided into 9 groups based on the band patterns. Moreover, isolates from one patient showed different band patterns. By RFLPs analysis, genetic heterogeneity seems to exist within the fimbrillin gene locus of P. gingivalis strains. Such genetic heterogeneity may reflect the previously reported difference of P. gingivalis fimbriae and moreover a single patient could be infected with more than one genotype of P. gingivalis.

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用限制性片段长度多态性分析牙龈卟啉单胞菌菌株在纤维蛋白基因位点上的异质性。
牙龈卟啉单胞菌(P. gingivalis)对牙周组织的粘附可能是牙周病发病的第一步,而菌毛被认为在这一过程中起着重要作用。然而,其大小、序列和抗原反应性的异质性已被报道。本研究的目的是利用RFLPs分析牙龈假单胞菌菌株在纤维蛋白基因位点上的异质性。选取47株牙龈卟啉单胞菌,其中5株为参考菌株。用亚硫酸酯修饰插入fimA381的质粒(pUC13Bg 12.1)作为探针。用限制性内切酶SacI和PstI酶切牙龈假单胞菌基因组DNA,进行电泳、转移和DNA探针杂交。除菌株W50外,所有菌株的Sac I酶解都产生一个主要条带,条带大小几乎相同(约2.5 kb)。Pst - 1型酶切结果显示出1 ~ 2个主要的谱带,根据谱带模式可分为9组。此外,一名患者的分离株显示不同的带型。通过RFLPs分析,牙龈假单胞菌菌株的纤维蛋白基因位点似乎存在遗传异质性。这种遗传异质性可能反映了先前报道的牙龈假单胞菌菌毛的差异,而且单个患者可能感染不止一种基因型的牙龈假单胞菌。
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