通过免疫荧光显微镜进行蛋白质细胞定位研究时对分枝杆菌包膜的通透处理。
Permeabilization of the mycobacterial envelope for protein cytolocalization studies by immunofluorescence microscopy.
作者信息
Cimino Mena, Alamo Lorenzo, Salazar Leiria
机构信息
Departamento de Biología Estructural, Instituto Venezolano de Investigaciones Científicas, Altos de Pipe, Estado Miranda, Venezuela.
出版信息
BMC Microbiol. 2006 Apr 18;6:35. doi: 10.1186/1471-2180-6-35.
BACKGROUND
The establishment of the cellular localization of proteins in M. tuberculosis will provide of valuable information for the identification of new drug/vaccine/diagnostic targets. Cytolocalization by inmunofluorescence microscopy has been limited in mycobacteria because to difficulties in effectively permeabilize it.
RESULTS
A treatment combining lysozyme with triton X-100 was found to be an effective permeabilization method of the mycobacterial envelope.
CONCLUSION
A rapid and simple permeabilization protocol has been successfully assessed in pure cultures of both Mycobacterium smegmatis and Mycobacterium tuberculosis H37Rv. This method can be successful used in the cytolocalization of proteins by immunolabeling.
背景
确定结核分枝杆菌中蛋白质的细胞定位将为鉴定新的药物/疫苗/诊断靶点提供有价值的信息。由于难以有效通透分枝杆菌，通过免疫荧光显微镜进行细胞定位在分枝杆菌中受到限制。
结果
发现溶菌酶与曲拉通X-100联合处理是一种有效的分枝杆菌包膜通透方法。
结论
已在耻垢分枝杆菌和结核分枝杆菌H37Rv的纯培养物中成功评估了一种快速简单的通透方案。该方法可成功用于通过免疫标记进行蛋白质的细胞定位。
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