| 重组Dsg3EC1-2及其嵌合毒素蛋白的原核表达及纯化 |
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| 引用本文： | 翟志芳,刁庆春,郝飞,钟白玉,唐书谦.重组Dsg3EC1-2及其嵌合毒素蛋白的原核表达及纯化[J].中国麻风皮肤病杂志,2005,21(3):169-171. |
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| 作者姓名： | 翟志芳 刁庆春 郝飞 钟白玉 唐书谦 |
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| 作者单位： | 1. 第三军医大学西南医院皮肤科,重庆,400038
2. 重庆市第一人民医院皮肤科,重庆,400011 |
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| 摘 要： | 目的：在大肠杆菌中高效表达并纯化重组天疱疮抗原桥粒芯糖蛋白3(Dsg3)的胞外结构域EC1-2片段及其与假单胞菌外毒素PE40的嵌合毒素Dsg3EC1-2PE40。方法：在大肠杆菌中对目的蛋白进行诱导表达，并用金属鏊合亲和层析法进行分离纯化，经Western Blot试验鉴定。结果：所得重组蛋白可以和寻常型天疱疮抗体IgG发生特异性结合，而与正常人IgG不发生反应，其纯化后纯度分别达85％及80%。结论：Dsg3EC1-2片段及其嵌合毒素Dsg3EC1-2PE40在原核细胞中得到成功表达，为天疱疮的治疗研究奠定了基础。
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| 关 键 词： | Dsg3 天疱疮 EC 嵌合 治疗 纯化 抗体IgG 桥粒芯糖蛋白 人IgG 正常人 |
Expression and purification of recombinant Dsg3EC1-2 and the chimeric toxin molecule in prokaryotic cells |
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| ZHAI Zhi-fang,DIAO Qing-chun,HAO Fei,et al..Expression and purification of recombinant Dsg3EC1-2 and the chimeric toxin molecule in prokaryotic cells[J].China Journal of Leprosy and Skin Diseases,2005,21(3):169-171. |
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| Authors: | ZHAI Zhi-fang DIAO Qing-chun HAO Fei |
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| Institution: | ZHAI Zhi-fang,DIAO Qing-chun,HAO Fei,et al. Department of Dermatology,Southwest Hospital,Third Military Medical University,Chongqing 400038 |
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| Abstract: | Objective: To effectively express and purify the recombinant Dsg3EC 1-2 and Dsg3EC 1-2PE40. Methods: The recombinant protein expressed by IPTG in Escherichia coli was induced, the proteins purified by the metal affinity chromatography and they were detected by Western blot. Results: The expressed proteins could conjugate with IgG from pemphigus vulguris patients but not do with IgG from the normal controls. The purity of Dsg3EC 1-2 and Dsg3EC 1-2PE40 was 85% and 80% respectively. Conclusion: The recombinant Dsg3EC 1-2 and Dsg3EC 1-2PE40 are expressed successfully in prokaryotic cells. |
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| Keywords: | pemphigus desmoglein chimeric toxin molecule metal affinity chromatography |
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