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Construction,Expression and Identification of Structural Gene for Porin I,the Major Outer Membrane Protein of Neisseria gonorrhoeae
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- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF DERMATOLOGY
- Issue:
- 6
- DOI:
- 10.3760/j.issn:0412-4030.2002.06.004
- Key Word:
- 奈瑟氏球菌,淋病;细菌外膜蛋白质类;重组融合蛋白质类
Abstract： 目的构建、克隆淋病奈瑟菌外膜PorinI(PI)蛋白基因,并进行表达、纯化和鉴定.方法用PCR法克隆淋病奈瑟菌外膜PI蛋白基因,再与pGEX-4T-2载体连接成表达重组体pGEX-4T-2/PI;经大肠杆菌P2392表达后,用SDS-PAGE、切胶、电洗脱回收等方法纯化GST-PI融合蛋白;然后用特异性淋球菌外膜PI蛋白的单克隆抗体进行斑点免疫层析试验鉴定该蛋白.结果成功地获取了pGEX-4T-2/PI表达重组体,经诱导表达后能获得高表达的GST-PI融合蛋白,其相对分子质量为60000;斑点免疫层析试验显示其为淋病奈瑟菌外膜PI蛋白特异性的.结论本研究将有利于进一步研究淋病奈瑟菌外膜PI蛋白的功能.
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