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1. 发明授权

US06329156B1 Method for screening inhibitors of the toxicity of Bacillus anthracis 失效
标题翻译：筛选炭疽芽孢杆菌毒性抑制剂的方法
公开(公告)号：US06329156B1
公开(公告)日：2001-12-11
申请号：US09273839
申请日：1999-03-22
申请人： Nick M. Cirino , Paul J. Jackson , Bruce E. Lehnert
发明人： Nick M. Cirino , Paul J. Jackson , Bruce E. Lehnert
IPC分类号： G01N33567
CPC分类号： C12Q1/18
摘要： The protective antigen (PA) of Bacillus anthracis is integral to the mechanism of anthrax poisoning. The cloning, expression and purification of a 32 kDa B. anthracis PA fragment (PA32) is described. This fragment has also been expressed as a fusion construct to stabilized green fluorescent protein (EGFP-PA32). Both proteins were capable of binding to specific cell surface receptors as determined by fluorescent microscopy and a flow cytometric assay. To confirm binding specificity in the flow cytometric assay, non-fluorescent PA83 or PA32 was used to competitively inhibit fluorescent EGFP-PA32 binding to cell receptors. This assay can be employed as a rapid screen for compounds which disrupts binding of PA to cells. Additionally, the high intracellular expression levels and ease of purification make this recombinant protein an attractive vaccine candidate or therapeutic treatment for anthrax poisoning.
摘要翻译：炭疽芽孢杆菌的保护性抗原（PA）是炭疽中毒机制的组成部分。描述了32kDa炭疽杆菌PA片段（PA32）的克隆，表达和纯化。该片段也被表达为稳定的绿色荧光蛋白（EGFP-PA32）的融合构建体。两种蛋白质都能够通过荧光显微镜和流式细胞术测定法测定特异性细胞表面受体。为了在流式细胞术检测中确认结合特异性，使用非荧光的PA83或PA32来竞争性地抑制荧光EGFP-PA32与细胞受体的结合。该测定法可用作快速筛选能破坏PA与细胞结合的化合物。此外，高细胞内表达水平和易于纯化使得该重组蛋白质成为有吸引力的候选药物或治疗炭疽中毒的治疗方法。

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