Gene Cloning of Glucosyltransferase Synthesizing Insoluble Glucan from Streptococcus mutans
| Project/Area Number |
60570881
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Functional basic dentistry
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| Research Institution | Nihon University |
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Principal Investigator |
ABIKO Yoshimitsu Nihon University School of Dentistry at Matsudo, Assistant Professor, 歯学部, 講師 (70050086)
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| Project Period (FY) |
1985 – 1986
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| Project Status |
Completed (Fiscal Year 1986)
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| Budget Amount *help |
¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1986: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1985: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Gene Cloning / Streptococcus mutans / Escherichia coli / Glucosyltransferase / Insoluble Glucan |
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| Research Abstract |
Gene banks of Streptococcus mutans B13N (serotype d) chromosomanl DNA in Escherichia coli were constructed by phage vector <lambda> L47.1 using <lambda> in vitro packaging technique. Phage gene banks were infected into E. coli, and glucosyltransferase synthesizing insoluble glucan (I-GTF) clones were screened by immuno-blot ELISA procedure using rabbit antiserum against I-GTF purified from S. mutans. Several clones had sucrose hydrolyzing activities. One clone designated as <lambda> MDSM39 was further studied. The I-GTF from <lambda> MDSM39 had molecular weight approximately 160 kdal by SDS-PAGE and HPLC gel filtration analysis, and exhibited a PI of approximately 3.8. This I-GTF activity was stimulated by the addition of soluble glucan synthesized GTF or dextran T10. The composition of insoluble glucan synthesized by <lambda> MDSM39 was mainly <alpha> -1,3 glucoside linkage. These data suggested that S. mutans I-GTF gene was successfully cloned and functionally expressed in E. coli. <lambda> MDSM39 clone will be a usefull material not only for enzymological and molecularbiological studies but also for development of vaccination against dental caries.
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Report
(1 results)
Research Products
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