Belardinelli Juan Manuel, Jackson Mary
Mycobacteria Research Laboratories, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523-1682, USA.
Mycobacteria Research Laboratories, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523-1682, USA.
Tuberculosis (Edinb). 2017 Jul;105:13-17. doi: 10.1016/j.tube.2017.04.001. Epub 2017 Apr 10.
The cell envelope-associated proteins of Mycobacterium species play critical functions in the physiology and pathogenicity of these microorganisms. Because the determination of their subcellular localization and transmembrane topology is often critical to the understanding of their function, we investigated whether the Green Fluorescent Protein (GFP) could be used as a reporter to probe protein localization and map the topology of inner membrane proteins directly in intact mycobacterial cells. To this end, two GFP-based mycobacterial reporter plasmids were engineered and their functionality validated using a variety of membrane-associated, exported and cytosolic proteins.
分枝杆菌属的细胞包膜相关蛋白在这些微生物的生理学和致病性中发挥着关键作用。由于确定它们的亚细胞定位和跨膜拓扑结构对于理解其功能通常至关重要，我们研究了绿色荧光蛋白（GFP）是否可用作报告基因，以直接在完整的分枝杆菌细胞中探测蛋白质定位并绘制内膜蛋白的拓扑结构。为此，构建了两个基于GFP的分枝杆菌报告质粒，并使用多种与膜相关、分泌和胞质蛋白验证了它们的功能。
