Bongaerts Roy J M, Hautefort Isabelle, Sidebotham Julie M, Hinton Jay C D
Institute of Food Research, Norwich Research Park, Norwich NR4 7UA, United Kingdom.
Methods Enzymol. 2002;358:43-66. doi: 10.1016/s0076-6879(02)58080-0.
To date, the majority of studies of bacterial gene expression have been carried out on large communities, as techniques for analysis of expression in individual cells have not been available. Recent developments now allow us to use reporter genes to monitor gene expression in individual bacterial cells. Conventional reporters are not suitable for studies of living single cells. However, variants of GFP have proved to be ideal for the study of development, cell biology, and pathogenesis and are now the reporters of choice for microbial studies. In combination with techniques such as DFI and IVET and the use of flow cytometry and advanced fluorescence microscopy, the latest generation of GFP reporters allows the investigation of gene expression in individual bacterial cells within particular environments. These studies promise to bring a new level of understanding to the fields of bacterial pathogenesis and environmental microbiology.
迄今为止,大多数关于细菌基因表达的研究都是在大型群落中进行的,因为尚无分析单个细胞中基因表达的技术。最近的进展使我们能够利用报告基因来监测单个细菌细胞中的基因表达。传统的报告基因不适用于活的单细胞研究。然而,绿色荧光蛋白(GFP)的变体已被证明是发育、细胞生物学和发病机制研究的理想选择,现在是微生物研究的首选报告基因。结合诸如差异荧光诱导(DFI)和体内表达技术(IVET)等技术以及流式细胞术和先进的荧光显微镜的使用,最新一代的GFP报告基因能够在特定环境中研究单个细菌细胞中的基因表达。这些研究有望为细菌发病机制和环境微生物学领域带来新的理解水平。