title: The identification of genes specific to Prevotella intermedia and Prevotella nigrescens using genomic subtractive hybridization.
abstract: Prevotella intermedia and Prevotella nigrescens, which are often isolated from periodontal sites, were once considered two different genotypes of P. intermedia. Although the genomic sequence of P. intermedia was determined recently, little is known about the genetic differences between P. intermedia and P. nigrescens. The subtractive hybridization technique is a powerful method for generating a set of DNA fragments differing between two closely related bacterial strains or species. We used subtractive hybridization to identify the DNA regions specific to P. intermedia ATCC 25611 and P. nigrescens ATCC 25261. Using this method, four P. intermedia ATCC 25611-specific and three P. nigrescens ATCC 25261-specific regions were determined. From the species-specific regions, insertion sequence (IS) elements were isolated for P. intermedia. IS elements play an important role in the pathogenicity of bacteria. For the P. intermedia-specific regions, the genes adenine-specific DNA-methyltransferase and 8-amino-7-oxononanoate synthase were isolated. The P. nigrescens-specific region contained a Flavobacterium psychrophilum SprA homologue, a cell-surface protein involved in gliding motility, Prevotella melaninogenica ATCC 25845 glutathione peroxide, and Porphyromonas gingivalis ATCC 33277 leucyl-tRNA synthetase. The results demonstrate that the subtractive hybridization technique was useful for distinguishing between the two closely related species. Furthermore, this technique will contribute to our understanding of the virulence of these species.
authors.affiliation: Division of Community Oral Health Science, Kyushu Dental College, Kitakyushu 803-8580, Japan.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Masakiyo
authors.forename: Akihiro
authors.initials: A
authors.lastname: Yoshida
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Shintani
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Ansai
authors.forename: Tadamichi
authors.initials: T
authors.lastname: Takehara
mesh_terms: D000217:Acyltransferases / Q000235:genetics*; D004251:DNA Transposable Elements / Q000235:genetics; D005798:Genes, Bacterial; D009693:Nucleic Acid Hybridization / Q000379:methods; D010518:Periodontitis / Q000382:microbiology; D018713:Prevotella intermedia / Q000235:genetics* / Q000472:pathogenicity; D045242:Prevotella nigrescens / Q000201:enzymology / Q000235:genetics*; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific) / Q000235:genetics; D013045:Species Specificity; D014774:Virulence / Q000235:genetics
chemical_list: D004251:DNA Transposable Elements; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific); D000217:Acyltransferases; C021929:8-amino-7-oxononanoate synthase