@article{oai:niigata-u.repo.nii.ac.jp:00025654,
 author = {中條, 和子 and 中澤, 太 and 星野, 悦郎},
 issue = {2},
 journal = {新潟歯学会雑誌, 新潟歯学会雑誌},
 month = {Dec},
 note = {Enterococcus Faecalis has been isolated frequently from human intestinal tracts. Also we have reported previously that these bacterial species has been occurred as predominant alkali resistant bacteria in human infected root canals. The aim of this study is to evaluate phylogenetic diversity of 16S rRNA of the human oral E. faecalis. In the present study, we used 9 strains of E. faecalis, ATCC 19433T, 4 strains isolated from human infected root canals and 4 strains isolated from human feces. From cells of these bacterial strains, genomic DNAs were extracted by using InstaGene Matrix and used as templates for amplification of 16S rDNA with universal primer sets in Polymerase Chain Reactions (PCR). For cycle sequence method of 16S rDNA sequence analysis, a Thermo Sequenase fluorescent labelled primer cycle sequencing kit was used with the 11 universal primer sets labeled with Cy-5, following the manufacture's protocol. Sequences of 16S rDNA were analyzed with a DNA sequencer. The segmented nucleotide sequences of 16S rDNA were connected using Seqman in LASERGENE computer program. Sequence similarity was analyzed by a clustal W method, which is programmed by Megalign in the LASERGENE computer program. 16S rDNA sequences of the E. faecalis reference strain (ATCC 19433T) was highly similar (98.4～100%) to those of the feces isolates and oral isolates, repectively. On the other hands, when the 16S rDNA sequences of oral isolates were compared to those of isolates from feces, these similarities were ranged widely and low (93.1～99.1%). These results of the present study has suggested that some of human oral E. faecalis may adapt to oral conditions and change genetically., Enterococcus faecalisはヒトの腸管細菌叢に広く生息している。我々はこれまで、本菌種が感染根管象牙質から検出されることや、強いアルカリ耐性を示すことを報告してきた｡今回は、生息の場の違いによる遺伝学的変異を明らかにする目的で、感染根管及び腸管分離E.faecalisについて、その16S rRNA geneの塩基配列を明らかにし、相互に比較検討した。E. faecalisの標準株ATCC 19433の他、感染根管象牙質、およびヒト糞便から得たE. faecalis分離株を用い、その培養菌体からInstaGene MatrixにてDNAを抽出した｡そのDNAをtemplateとし、universal primersを用いてPCR法にて16S rDNAを増幅後、精製16S rDNA、 Thermo Sequenase Labelled Primer Cycle Sequencing Kit with 7-deazadGTPと11種類のprimerにてPCRLし、ALF expressを使ってその塩基配列を明らかにした。更にsequence解析プログラムLASERGENEを用いて相互のsequence similarityを明らかにし、遺伝学的相互関係を検討した｡その結果、根管由来と腸管由来の菌株それぞれと、標準株との比較では著しく高い相同性(98.4-100%)を示したのに対し、根管由来と腸管由来の菌株間の相同性は93.1-99.1％であった。 これらの結果から、根管由来と腸管由来のE.Faecalisにおいでは、生息の場の違いと共に菌株間にも遺伝学的差異が認められた。},
 pages = {17--23},
 title = {ヒト口腔由来と腸管由来のEnterococcus faecalisにおける遺伝学的検討},
 volume = {33},
 year = {2003}
}