Gunning A P, Bongaerts R J M, Kirby A R, Hinton J C D, Morris V J
Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, U.K.
J Microsc. 2005 Apr;218(Pt 1):46-51. doi: 10.1111/j.1365-2818.2005.01464.x.
IcsA is an autotransporter protein that plays a role in the virulence of Shigella bacteria. We have examined the cellular localization of a fusion of an IcsA fragment to the green fluorescent protein (GFP) expressed in Escherichia coli using a dual epifluorescence and scanning near-field optical microscope. By combining the data obtained from far-field with near-field microscopy of the same sample, discrimination between surface-bound fusion proteins and fusion proteins located in the cellular cytoplasm becomes possible. Furthermore, and for the first time, the inherent advantages in resolution of the near-field images provides highly specific details of the location of a GFP fusion protein on a bacterial cell surface.
IcsA是一种自转运蛋白，在志贺氏菌的毒力中发挥作用。我们使用双落射荧光和扫描近场光学显微镜检查了在大肠杆菌中表达的IcsA片段与绿色荧光蛋白（GFP）融合体的细胞定位。通过将从远场获得的数据与同一样本的近场显微镜数据相结合，就能够区分表面结合的融合蛋白和位于细胞质中的融合蛋白。此外，近场图像分辨率的固有优势首次提供了GFP融合蛋白在细菌细胞表面位置的高度特异性细节。
