摘要
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。
PURPOSE： To prepare and identify polyclonal antibody of Streptococcus salivarius 57.I urease. METHODS： The biggest structural subunit of Streptococcus salivarius 57.I nrease, UreC, was obtained by gene clone, IPTG-induced expression, and purification through affinity chromatography. Anti-sera and polyelonal antibody were raised by immunizing rabbits with purified UreC. Western blot was utilized to detect the specific combination of polyclonal antibody with UreC. RESULTS： Purified UreC protein was prepared and used as antigen to immunize rabbits. Polyclonal antibody was obtained, and Western blot displayed a specific band of the polyclonal antibody with UreC about 62 kD as anticipated. CONCLUSIONS： Polyclonal antibody against Streptococcus salivarius 57.I urease is obtained, which provides an important tool to explore the function of urease and its relationship with dental caries.
出处
《上海口腔医学》
CAS
CSCD
2016年第6期678-681,共4页
Shanghai Journal of Stomatology
基金
上海市自然科学基金(13ZR1436200)
上海市卫生与计划生育委员会重点项目(20134028)
关键词
唾液链球菌
尿素酶
抗体
Streptococcus salivarius
Urease
Antibody
