Electrotransformation of streptococcus with lacotococci plasmid and investigation of the factor of dental caries by oral microorganisms
| Project/Area Number |
11672036
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
矯正・小児・社会系歯学
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| Research Institution | TOHOKU UNIVERSITY |
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Principal Investigator |
HATA Shinji TOHOKU UNIVERSITY, GRADUATE SCHOOL OF DENTISTRY, DEPARTMENT OF LIFELONG ORAL HEALTH SCIENCES, DIVISION OF PEDIATRIC DENTISTRY, LECTURER, 大学院・歯学研究科, 講師 (20208507)
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| Co-Investigator(Kenkyū-buntansha) |
HATA Hiroko TOHOKU UNIVERSITY, SCHOOL OF DENTISTRY, RESEARCH ASSISTANT, 歯学部・附属病院, 助手 (20180630)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2000: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1999: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | PLASMID / LACTOCOCCUS / ORAL STREPTOCOCCI / β-GALACTOSUGARS / プラスミドベクター / ストレプトコッカス / 耐酸性因子 |
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| Research Abstract |
1. Extraction pGB301 from Lactococcus lactis JK301
pGB301 was extracted by means of alkali-SDS and ultracentrifusion. A single band was confirmed on agarosen gel electrophoresis.
2. Electrotransformation of Streptococcus milleli 10707, S.sanguis ATCC 35105 and S.gordonii Challis and its efficiency
The strains of the microorganisms were cultured in Todd-Hewitt broth sulpplemented with 5% horse serum. The microorganisms grew on plate with Erythromycin by the transformation. Its efficiency was ranged form 1.5×10^4 to 5.5×10^4. The effciency increased five times by electroporation. It was most effective whhen the microorganisms were harvested at 0.1 of optical density of the broth.
3. Construction of low-pH sensitve mutansts of S.milleli 10707.
Low-pH sensitve mutansts of S.milleli 10707 were constructed with ethyl methasulfonete. The parent strain grew in the broth of pH 4.2. On the other hand the mutant did not grow in the broth of pH 4.6. Its transformation efficiency was 1.4 ×10^3 CFU/μg.
4. The most effective condition for electrotransformation
The most effective condition for electrotransformation was obtained when peak voltage was 2.5 kV, capacitance was 25 F, pulse controll was 200Ω and harvest OD was 0.5.
5. Cariogenic factos of β-galacto-sugars by oral microorganisms and the activity of the enzymes involved
Mutans streptococci did not utilize β-galactosyllactose and lactosylfructoside.
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Report
(3 results)
Research Products
(3 results)
