1999 Fiscal Year Final Research Report Summary
Replacement Therapy for Dental Caries by Antibody Secretion System.
| Project/Area Number |
10671719
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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 |
Morphological basic dentistry
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| Research Institution | Nihon University |
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Principal Investigator |
HAYAKAWA Mitsuo Nihon Unversity Schol of Dentistry at Matsudo Lecturer, 松戸歯学部, 教授 (10112955)
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| Project Period (FY) |
1998 – 1999
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| Keywords | dental caries / replacement therapy / secretive vector / insoluble protein / electro-osmosis / electropholetic purification |
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| Research Abstract |
In the oral cavity, bacterial pathogens are often associated with one of major oral disease, dental caries. The colonization and accumulation of cariogenic oral streptococci on the tooth surafce have been associated with the ability to synthesize water insoluble glucans from sucrose by glucosyltransferase (gtfs). In this study, we attempted to develop an advanced replacement therapy against dental caries utilizing secretive antibody to gtf I by biotechnological approaches. Single chain variable fragments (ScFv) of antibody genes were cloned from the hybridoma mRNA and expressed into E.coli using RT-PCR technique (Shibata et al. Infect Immun 1987). A novel transformation technique, resident plasmid integration (Siroza et al. Gene, 1998), was used to clone the gene coding of (ScFv) antibody to gtf. I, which inhibit a gtf I synthetic activity, from E.coli. Into S.gordonii and S.brebis. The gene product was successfully expressed and extracellularlly secreted in S.brebis as an insoluble form. Then we constructed and developed an improved electro-osmotic elution system for preparative sodium dodecyl sulfate poly acryl amide gel electrophoresis based on the idea of Tan et al. (1988) in a large scale. In this elution system, a secreted protein, ScFv polypeptide was purified in size and recovered with a high concentration and in a high yield.
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