Phillips G J
Department of Microbiology, 207 Science I Building, Iowa State University, Ames, IA 50011, USA.
FEMS Microbiol Lett. 2001 Oct 16;204(1):9-18. doi: 10.1111/j.1574-6968.2001.tb10854.x.
Use of the green fluorescent protein (GFP) of Aequorea victoria as a reporter for protein and DNA localization has provided sensitive, new approaches for studying the organization of the bacterial cell, leading to new insights into diverse cellular processes. GFP has many characteristics that make it useful for localization studies in bacteria, primarily its ability to fluoresce when fused to target polypeptides without the addition of exogenously added substrates. As an alternative to immunofluorescence microscopy, the expression of gfp gene fusions has been used to probe the function of cellular components fundamental for DNA replication, translation, protein export, and signal transduction, that heretofore have been difficult to study in living cells. Moreover, protein and DNA localization can now be monitored in real time, revealing that several proteins important for cell division, development and sporulation are dynamically localized throughout the cell cycle. The use of additional GFP variants that permit the labeling of multiple components within the same cell, and the use of GFP for genetic screens, should continue to make this a valuable tool for addressing complex questions about the bacterial cell.
利用维多利亚多管水母的绿色荧光蛋白（GFP）作为蛋白质和DNA定位的报告基因，为研究细菌细胞的组织结构提供了灵敏的新方法，从而对多种细胞过程有了新的认识。GFP具有许多使其适用于细菌定位研究的特性，主要是它在与靶多肽融合时能够发出荧光，而无需添加外源底物。作为免疫荧光显微镜的替代方法，gfp基因融合的表达已被用于探究对DNA复制、翻译、蛋白质输出和信号转导至关重要的细胞成分的功能，而这些成分迄今为止在活细胞中难以研究。此外，现在可以实时监测蛋白质和DNA的定位，这表明几种对细胞分裂、发育和孢子形成重要的蛋白质在整个细胞周期中动态定位。使用能够标记同一细胞内多个成分的其他GFP变体，以及将GFP用于遗传筛选，应该会继续使它成为解决有关细菌细胞复杂问题的有价值工具。
