论文部分内容阅读
目的：变形链球菌表面蛋白抗原(surface protein antigen, PAc)功能区A区（简称PAc/A）的主要功能是参与前期细菌与牙齿的粘附作用，是变异链球菌发挥致病作用的基础。目的：通过基因克隆及表达的基因工程手段制备变形链球菌表面蛋白抗原A区的特异性多克隆抗体。方法：获得变形链球菌表面蛋白抗原功能区A区的基因克隆，构建表达质粒和转染大肠杆菌，IPTG诱导表达，亲和层析纯化并定量检测，利用纯化后的抗原蛋白免疫新西兰大白兔制备多克隆抗体，Western blot鉴定免疫活性和特异性。结果：Western blot法表明多克隆抗体能够识别PAc蛋白分子，并且与PA c特异性结合，为基因疫苗与蛋白质免疫相结合的龋病防治新策略奠定基础。“,”A function in the surface protein antigen(PAc/A)of Streptococcus mutans is to take part in the adhesion of the bacteria to the teeth, and it is the basis of the pathogenic role of Streptococcus mutants. PURPOSE: The purpose of this study was to prepare and identify polyclonal antibody of PAc/A by the method of cloning and expression of gene. METHODS: PAc/A was obtained by gene cloning, induced expression by IPTG, and purified by affinity chromatography. Anti-sera and polyclonal antibody were raised by immunizing rabbit with purified PAc/A. Western Blot was utilized to detect the specific combination of polyclonal antibody with PAc/A. RESULTS: The method of Western blot showed that the polyclonal antibody can recognize protein of PAc/A, and specific combined with the PAc/A. It wil lay the foundation for studying the new strategy for the treatment of dental caries.