唾液链球菌57.I 尿素酶抗体的制备和鉴定

上海口腔医学 ›› 2016, Vol. 25 ›› Issue (6): 678-681.

唾液链球菌57.I 尿素酶抗体的制备和鉴定

王艳¹, 陶丹英², 冯希平²

1.上海市口腔病防治院·上海市口腔医院口腔预防处,上海 200001;;
2.上海交通大学医学院附属第九人民医院·口腔医学院口腔预防科, 上海市口腔医学重点实验室,上海 200011 ;

收稿日期:2016-05-11 出版日期:2016-12-25 发布日期:2016-12-29
通讯作者: 冯希平,E-mail: fxiping1808@qq.com
作者简介:王艳（1978-）,女,博士,副主任医师,E-mail: serene2000@163.com
基金资助:
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）

Preparation and identification of polyclonal antibody of Streptococcus salivarius 57.I urease

WANG Yan¹, TAO Dan-ying², FENG Xi-ping²

1.Department of Preventive Dentistry, Shanghai Stomatological Hospital. Shanghai 200001;
2.Department of Preventive Dentistry, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine; Shanghai Key Laboratory of Stomatology. Shanghai 200011, China

Received:2016-05-11 Online:2016-12-25 Published:2016-12-29

摘要/Abstract

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。

关键词: 唾液链球菌, 尿素酶, 抗体

Abstract: PURPOSE: To prepare and identify polyclonal antibody of Streptococcus salivarius 57.I urease. METHODS: The biggest structural subunit of Streptococcus salivarius 57.I urease, UreC, was obtained by gene clone, IPTG-induced expression, and purification through affinity chromatography. Anti-sera and polyclonal 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.

Key words: Streptococcus salivarius, Urease, Antibody

中图分类号:

R780.2

王艳, 陶丹英, 冯希平. 唾液链球菌57.I 尿素酶抗体的制备和鉴定[J]. 上海口腔医学, 2016, 25(6): 678-681.

WANG Yan, TAO Dan-ying, FENG Xi-ping. Preparation and identification of polyclonal antibody of Streptococcus salivarius 57.I urease[J]. Shanghai Journal of Stomatology, 2016, 25(6): 678-681.

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