| Project/Area Number |
05454493
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Morphological basic dentistry
|
|---|
| Research Institution | Kanagawa Dental College |
|---|
Principal Investigator |
UMEMOTO Toshio Kanagawa Dental College, Dept.of Oral Microbiology, Professor, 歯学部, 教授 (20067036)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Yusuke Kanagawa Dental College, Dept.of Oral Microbiology, Assistant, 歯学部, 助手 (20267511)
HAMADA Nobushiro Kanagawa Dental College, Dept.of Oral Microbiology, Assistant, 歯学部, 助手 (20247315)
YOSHIMOTO Hisashi Kanagawa Dental College, Dept.of Oral Microbiology, Lecturer, 歯学部, 講師 (60084787)
|
|---|
| Project Period (FY) |
1993 – 1995
|
| Project Status |
Completed (Fiscal Year 1995)
|
| Budget Amount *help |
¥5,700,000 (Direct Cost: ¥5,700,000)
Fiscal Year 1995: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1994: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1993: ¥3,300,000 (Direct Cost: ¥3,300,000)
|
|---|
| Keywords | Periodontpathic bacteria / P.gingivalis / Physical map / NotI fragment / Genome size / Gene map / Virulent gene / Pulsed-field gel electrophoresis / 歯周病原細菌 / Porphyromonas gingivalis / 染色体DNA / 制限酵素地図 / 遺伝子地図 |
|---|
| Research Abstract |
Genetic approaches have been recently introduced to analyze virulence factors of Porphyromonas gingivalis. Several genes potentially involved in their pathogenicity related with periodontal disease. However, little is known of the over all genetic organization of this bacterium. Recently, pulsed-field gel electrophoresis was developed to separate large DNA fragment. Physical maps of several bacterial genome have been constructed by using pulsed-field gel electrophoresis. In this study, a physical and gene map of genome of P.gingivalis ATCC 33277 constructed by Southern blot analysis of NotI restriction fragments separated by pulsed-field gel electrophoresis. We firstly examined polymorphism of the patterns of the NotI-digested chromosomal DNA fragments of the strains ATCC 33277,381, ATCC 53977, W50, and W83. All these strains showed completely different patterns, though only those of W50 and W83 were very similar.Then we tried to construct a NotI map of the chromosomal DNA of P.gingivalis ATCC 33277. The restriction enzyme NotI produced at least 10 fragments, which were ranging between kilobases of 30 and 690. These sizes were determined by pulsed-field gel electrophoresis. Thus, the total size of the chromosome was estimated to be about 2600 kb. Subsequently, Southern hybridization analysis of the NotI-digested chromosome was performed with some DNA probes, such as the fimbrillin gene (fimA), the superoxide dismutase gene (sod) and one of the genes of trypsin-like protease. The probe for fimA gene hybridized to the largest fragment and that for sod to the fifth. The probe for the trypsin gene hybridized to the sixth fragment from the top. These results suggested that there was no close linkage among these three genes examined.
|
