Tachibana-Ono M, Yoshida A, Kataoka S, Ansai T, Shintani Y, Takahashi Y, Toyoshima K, Takehara T
Division of Community Oral Health Science, Kyushu Dental College, Kokurakita-ku, Kitakyushu, Japan.
Oral Microbiol Immunol. 2008 Feb;23(1):84-7. doi: 10.1111/j.1399-302X.2007.00396.x.
BACKGROUND/AIMS: Porphyromonas gingivalis, a major etiological organism implicated in periodontal disease, can be classified into virulent and avirulent strains. Our aim was to identify a gene for the virulence of P. gingivalis.
The subtractive hybridization technique was employed to identify the genes specific to P. gingivalis W83, a virulent strain. In this study, P. gingivalis W83 was used as the tester strain, and P. gingivalis ATCC 33277 was the driver strain. The prevalence of W83-specific genes was determined by Southern blot analysis of several P. gingivalis strains.
We obtained 575 colonies using the subtractive hybridization technique. From among these, 26 DNA fragments were subjected to a homology search using the BLAST program. Compared with strain ATCC 33277, strain W83 contained 12 unique clones. The specificities of the isolated DNA fragments were analyzed among four P. gingivalis strains by Southern blot analysis. Five genes showed specificity for strain W83 compared with strain ATCC 33277. All five genes were also identified in strain W50.
The subtractive hybridization technique was effective in screening the two strains for specific DNA sequences, some of which might be responsible for determining virulence. The results suggested that several genes specific to strain W83 were associated with its virulence. Further analysis of these DNA fragments will provide important information on the pathogenesis of virulent P. gingivalis strains.
背景/目的：牙龈卟啉单胞菌是导致牙周疾病的一种主要病原体，可分为有毒力菌株和无毒力菌株。我们的目的是鉴定牙龈卟啉单胞菌毒力相关基因。
采用消减杂交技术鉴定有毒力的牙龈卟啉单胞菌W83菌株的特异性基因。在本研究中，牙龈卟啉单胞菌W83用作检测菌株，牙龈卟啉单胞菌ATCC 33277用作驱动菌株。通过对几种牙龈卟啉单胞菌菌株进行Southern印迹分析来确定W83特异性基因的普遍性。
利用消减杂交技术我们获得了575个菌落。其中，26个DNA片段使用BLAST程序进行同源性搜索。与ATCC 33277菌株相比，W83菌株含有12个独特的克隆。通过Southern印迹分析在4种牙龈卟啉单胞菌菌株中分析了分离的DNA片段的特异性。与ATCC 33277菌株相比，5个基因对W83菌株具有特异性。在W50菌株中也鉴定出了所有这5个基因。
消减杂交技术有效地筛选了这两种菌株的特异性DNA序列，其中一些可能与毒力的决定有关。结果表明，W83菌株的几个特异性基因与其毒力相关。对这些DNA片段的进一步分析将为有毒力的牙龈卟啉单胞菌菌株的发病机制提供重要信息。
