INHIBITION OF HUMAN CARIOGENIC STREPTOCOCCI

对人类致龋链球菌的抑制

• 批准号: 3218862
• 负责人: GERALD D SHOCKMAN
• 金额: $ 14.15万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 1975
• 资助国家: 美国
• 起止时间: 1975-01-01 至 1989-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3218862
• 关键词: Streptococcus mutans; antibacterial agents; cell wall; dental caries; dental caries inhibitor; dental plaque; electron microscopy; immunochemistry; microorganism culture; monoclonal antibody; oral bacteria; polymers; radiotracer; scintillation counter

Using well characterized, agglutinating, monoclonal antibodies directed against various surface constituents, non-agglutinating mutants of Streptococcus mutans GS-5 will be isolated. Among these will be mutants defective in a range of surface constituents including serotype c antigen, lipoteichoic acid, glucosyl transferase(s), and other surface antigens. In addition, attempts will be made to isolate GS-5 organisms defective in lactic dehydrogenase. Plasmids from an already available gene bank (generated in E. coli and consisting of GS-5 DNA inserts into a chimeric plasmid capable of replication in E. coli and in Streptococci) will be used to transform each deficient GS-5 mutant to normal surface function. The complemented function will be detected, enriched and selected by agglutination using the previously mentioned monoclonal antibodies. An alternate protocol, using transposon inactivation to generate both mutants and cloned revertants, is provided and is considered especially useful for the isolation of lactic dehydrogenese deficient organisms. The project is designed to provide well characterized, truly isogneic pairs of GS-5 derivatives defective in various properties thought to be associated with cariogenicity. In vitro assays for virulence will include studies of interactions between hydrophobic constitutents, lectin-ligand interactions, formation of adhesive polysaccharides, and bacterial-bacterial interactions. We also plan to establish collaborative studies with other investigators for in vivo animal studies of these isogenic pairs.

使用良好的特征，凝集的单克隆抗体定向 针对各种表面成分，非凝集突变体 将分离出链球菌突变体GS-5。 其中将是突变体 在包括血清型C抗原在内的一系列表面成分中有缺陷， 脂肪酸，葡萄糖基转移酶（S）和其他表面抗原。 在 此外，将尝试隔离GS-5生物有缺陷的GS-5生物 乳酸脱氢酶。 来自已经可用的基因库的质粒 （以大肠杆菌的形式产生，由GS-5 DNA插入到嵌合 将使用能够在大肠杆菌和链球菌中复制的质粒） 将每个缺陷的GS-5突变体转换为正常表面功能。 这 将检测，丰富和选择补充功能 使用先前提到的单克隆抗体的凝集。 一个 替代方案，使用转座子灭活来生成两个突变体 和克隆的恢复物，被提供，被认为是特别有用的 乳酸脱氢酶缺乏生物的分离。 该项目是 设计旨在提供良好的GS-5的特征性，真正的异源对 被认为与之相关的各种特性中有缺陷的衍生物 致癌性。 毒力的体外测定法包括 疏水组成部分，凝集素 - 配体相互作用之间的相互作用， 形成粘合多糖和细菌细菌的形成 互动。 我们还计划与其他 研究这些同源性对体内动物研究的研究者。