Replacement Therapy for Dental Caries by Antibody Secretion System.

在口腔中，细菌性病原体往往与龋齿这一口腔主要疾病有关。龋齿性口腔链球菌在牙齿表面的定植和积累与通过葡萄糖基转移酶（gtfs）从蔗糖合成水不溶性葡聚糖的能力有关。在这项研究中，我们试图通过生物技术方法开发一种先进的替代疗法，利用gtf I的隐秘抗体来治疗龋齿。单链可变片段（ScFv）抗体基因从杂杂瘤mRNA中克隆出来，利用RT-PCR技术在大肠杆菌中表达（Shibata et al.）。感染免疫，1987)。一种新的转化技术，驻留质粒整合（Siroza等）。Gene, 1998)，用于克隆（ScFv）抗gtf的编码基因。I，抑制gtf I合成活性，来自大肠杆菌。分为S.gordonii和S.brebis。该基因产物成功表达，并以不溶性形式在s.b bretis细胞外分泌。然后，我们基于Tan et al.（1988）的思想，大规模构建并开发了一种改进的制备十二烷基硫酸钠聚丙烯酰胺凝胶电泳的电渗透洗脱系统。在该洗脱系统中，一种分泌蛋白，ScFv多肽被纯化，并以高浓度和高产率回收。

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.