Identification by subtractive hybridization of a novel insertion sequence specific for virulent strains of Porphyromonas gingivalis

Koichi Sawada, Susumu Kokeguchi, Hiroshi Hongyo, Satoko Sawada, Manabu Miyamoto, Hiroshi Maeda, Fusanori Nishimura, Shogo Takashiba, Yoji Murayama

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Subtractive hybridization was employed to isolate specific genes from virulent Porphyromonas gingivalis strains that are possibly related to abscess formation. The genomic DNA from the virulent strain P. gingivalis W83 was subtracted with DNA from the avirulent strain ATCC 33277. Three clones unique to strain W83 were isolated and sequenced. The cloned DNA fragments were 885, 369, and 132 bp and had slight homology with only Bacillus stearothermophilus IS5377, which is a putative transposase. The regions flanking the cloned DNA fragments were isolated and sequenced, and the gene structure around the clones was revealed. These three clones were located side-by-side in a gene reported as an outer membrane protein. The three clones interrupt the open reading frame of the outer membrane protein gene. This inserted DNA, consisting of three isolated clones, was designated IS1598, which was 1,396 bp (i.e., a 1,158-bp open reading frame) in length and was flanked by 16-bp terminal inverted repeats and a 9-bp duplicated target sequence. IS1598 was detected in P. gingivalis W83, W50, and FDC 381 by Southern hybridization. All three P. gingivalis strains have been shown to possess abscess-forming ability in animal models. However, IS1598 was not detected in avirulent strains of P. gingivalis, including ATCC 33277. The IS1598 may interrupt the synthesis of the outer membrane protein, resulting in changes in the structure of the bacterial outer membrane. The IS1598 isolated in this study is a novel insertion element which might be a specific marker for virulent P. gingivalis strains.

Original languageEnglish
Pages (from-to)5621-5625
Number of pages5
JournalInfection and Immunity
Volume67
Issue number11
Publication statusPublished - Nov 1999
Externally publishedYes

Fingerprint

Porphyromonas gingivalis
Insertional Mutagenesis
Clone Cells
DNA
Membrane Proteins
Abscess
Open Reading Frames
Genes
Transposases
Bacterial Structures
Geobacillus stearothermophilus
Terminal Repeat Sequences
Subtractive Hybridization Techniques
Animal Models
Membranes

All Science Journal Classification (ASJC) codes

  • Immunology

Cite this

Sawada, K., Kokeguchi, S., Hongyo, H., Sawada, S., Miyamoto, M., Maeda, H., ... Murayama, Y. (1999). Identification by subtractive hybridization of a novel insertion sequence specific for virulent strains of Porphyromonas gingivalis. Infection and Immunity, 67(11), 5621-5625.

Identification by subtractive hybridization of a novel insertion sequence specific for virulent strains of Porphyromonas gingivalis. / Sawada, Koichi; Kokeguchi, Susumu; Hongyo, Hiroshi; Sawada, Satoko; Miyamoto, Manabu; Maeda, Hiroshi; Nishimura, Fusanori; Takashiba, Shogo; Murayama, Yoji.

In: Infection and Immunity, Vol. 67, No. 11, 11.1999, p. 5621-5625.

Research output: Contribution to journalArticle

Sawada, K, Kokeguchi, S, Hongyo, H, Sawada, S, Miyamoto, M, Maeda, H, Nishimura, F, Takashiba, S & Murayama, Y 1999, 'Identification by subtractive hybridization of a novel insertion sequence specific for virulent strains of Porphyromonas gingivalis', Infection and Immunity, vol. 67, no. 11, pp. 5621-5625.
Sawada, Koichi ; Kokeguchi, Susumu ; Hongyo, Hiroshi ; Sawada, Satoko ; Miyamoto, Manabu ; Maeda, Hiroshi ; Nishimura, Fusanori ; Takashiba, Shogo ; Murayama, Yoji. / Identification by subtractive hybridization of a novel insertion sequence specific for virulent strains of Porphyromonas gingivalis. In: Infection and Immunity. 1999 ; Vol. 67, No. 11. pp. 5621-5625.
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