| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1999: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1998: ¥1,200,000 (Direct Cost: ¥1,200,000)
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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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