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目的 :利用基因克隆和表达的多肽制备唾液链球菌57.I尿素酶的特异性多克隆抗体。方法 :获得唾液链球菌57.I尿素酶最大的结构亚基ureC的克隆,IPTG诱导表达、纯化并定量,用其免疫新西兰大白兔,获得抗血清和多克隆抗体,采用Western印迹法鉴定其特异性。结果:获得了纯化UreC蛋白,以其作为抗原免疫,制备出多克隆抗体。经Western印迹试验表明,该多克隆抗体可识别UreC目的蛋白条带,其相对分子量约为62 kD,与预测分子量相符。结论:所制备的多克隆抗体可识别UreC蛋白,能够与唾液链球菌57.I尿素酶抗原特异性结合,为深入研究尿素酶的功能及其与龋病的关系提供了有力工具。
OBJECTIVE: To prepare specific polyclonal antibodies to Streptococcus salivarius 57.I urease using the gene cloned and expressed polypeptide. Methods: Clone of urease, the largest structural unit of Streptococcus salivarius 57.I urease, was induced by IPTG, purified and quantified. New Zealand white rabbits were immunized with it to obtain antiserum and polyclonal antibody. The specificity was identified by Western blotting Sex. Results: Purified UreC protein was obtained and used as antigen to prepare polyclonal antibody. Western blotting experiments showed that the polyclonal antibody can recognize the UreC target protein band, the relative molecular weight of about 62 kD, consistent with the predicted molecular weight. CONCLUSION: The prepared polyclonal antibody can recognize UreC protein and bind specifically with Streptococcus salivarius 57.I urease antigen, which provides a powerful tool for further study on the function of urease and its relationship with caries.