Kocaoglu Ozden, Carlson Erin E
Department of Molecular and Cellular Biochemistry, Indiana University, Bloomington, Indiana, USA.
Department of Chemistry, Indiana University, Bloomington, Indiana, USA.
Nat Chem Biol. 2016 Jun 17;12(7):472-8. doi: 10.1038/nchembio.2109.
Fluorescence microscopy is an essential tool for the exploration of cell growth, division, transcription and translation in eukaryotes and prokaryotes alike. Despite the rapid development of techniques to study bacteria, the size of these organisms (1-10 μm) and their robust and largely impenetrable cell envelope present major challenges in imaging experiments. Fusion-based strategies, such as attachment of the protein of interest to a fluorescent protein or epitope tag, are by far the most common means for examining protein localization and expression in prokaryotes. While valuable, the use of genetically encoded tags can result in mislocalization or altered activity of the desired protein, does not provide a readout of the catalytic state of enzymes and cannot enable visualization of many other important cellular components, such as peptidoglycan, lipids, nucleic acids or glycans. Here, we highlight the use of biomolecule-specific small-molecule probes for imaging in bacteria.
荧光显微镜是探索真核生物和原核生物细胞生长、分裂、转录和翻译的重要工具。尽管研究细菌的技术发展迅速，但这些生物体的大小（1-10μm）以及它们坚固且大多难以穿透的细胞膜在成像实验中带来了重大挑战。基于融合的策略，例如将感兴趣的蛋白质与荧光蛋白或表位标签连接，是目前在原核生物中检测蛋白质定位和表达最常用的方法。虽然很有价值，但使用基因编码标签可能会导致所需蛋白质的定位错误或活性改变，无法提供酶催化状态的读数，也无法实现对许多其他重要细胞成分的可视化，如肽聚糖、脂质、核酸或聚糖。在这里，我们重点介绍生物分子特异性小分子探针在细菌成像中的应用。
