国际口腔医学杂志 ›› 2015, Vol. 42 ›› Issue (6): 655-658.doi: 10.7518/gjkq.2015.06.010

• 论著 • 上一篇    下一篇

牙龈卟啉单胞菌菌毛蛋白的重组及表达

肖莉1 林玉祥2 葛颂3   

  1. 1.合肥市口腔医院牙周科 合肥 230000; 2.厦门市口腔医院牙周科 厦门 361003; 3.遵义医学院附属口腔医院 遵义 563003
  • 收稿日期:2015-03-10 修回日期:2015-08-20 出版日期:2015-11-01 发布日期:2015-11-01
  • 通讯作者: 葛颂,教授,博士,Email:gesong007b@163.com
  • 作者简介:肖莉,主治医师,硕士,Email:150811519@qq.com
  • 基金资助:

    国家自然科学基金(30760271,81260168);贵州省优秀青年科技人才培养对象专项资金(黔科合人字[2011]32号)

Recombination and expression of fimbriae protein for Porphyromonas gingivalis

Xiao Li1, Lin Yuxiang2, Ge Song3.   

  1. 1. Dept. of Periodontics, Hefei Stomatological Hospital, Hefei 230000, China; 2. Dept. of Periodontics, Xiamen Stomatological Hospital, Xiamen 361003, China; 3. Stomatological Hospital Affiliated to Zunyi Medical College, Zunyi 563003, China
  • Received:2015-03-10 Revised:2015-08-20 Online:2015-11-01 Published:2015-11-01

摘要:

目的 克隆Ⅱ fimA型牙龈卟啉单胞菌菌毛fimA基因并使其在结肠埃希菌中正确表达。方法 利用聚合酶链反应技术克隆Ⅱ型 fimA基因,经酶切和测序验证后构建原核表达质粒pET-FimA,使其在结肠埃希菌中表达,十二烷基硫酸钠聚丙烯酰胺凝胶电泳和Western blot鉴定表达产物。结果 克隆基因测序结果与GeneBank数据库中的序列呈现95%同源性,成功构建 fimA基因原核表达载体pET-FimA。结论 高效表达出Ⅱ fimA型重组菌毛蛋白,并获得较高纯度,且具有免疫原性和免疫反应性。

关键词: Ⅱ fimA型牙龈卟啉单胞菌, 重组菌毛, 蛋白表达

Abstract:

Objective This study aimed to clone the type Ⅱ fimA gene of Porphyromonas gingivalis(P. gingivalis) and detect its expression in Escherichia coli(E. coli). Methods The type Ⅱ fimA gene was obtained by polymerase chain reaction(PCR) from the genome of P. gingivalis to construct a prokaryotic expression plasmid pET-FimA. pET-FimA was transformed into E. coli BL21(DE3) competent cells, and the recombination protein was characterized via sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) and Western blot analysis. Results DNA sequencing showed that the fragment was 95% consistent with that of published literature. We successfully constructed prokaryotic expression plasmid pET-FimA. Conclusion The plasmid efficiently expressed the recombinant fimbriae protein of the type ⅡfimA gene of P. gingivalis. A high purity of protein FimA was obtained. FimA has immunogenicity and immune reactivity.

Key words: type Ⅱ fimA gene of Porphyromonas gingivalis, recombinant fimbriae protein, protein expression

中图分类号: 

  • R 782