Samalova Marketa, Fricker Mark, Moore Ian
Department of Plant Sciences, University of Oxford, Oxford OX1 3RB, United Kingdom.
Methods Cell Biol. 2008;85:353-80. doi: 10.1016/S0091-679X(08)85015-7.
Fluorescent proteins have had a great impact on the way in which plant membrane traffic is studied. Here we review the uses to which these molecules have been put in this field of research and discuss the advantages and pitfalls of particular fluorescent protein derivatives in various applications and plant species. We discuss in detail the need for quantitative estimates of expression level and the potential of fluorescent proteins for quantitative assays of biosynthetic membrane traffic. Detailed descriptions and protocols are provided for the use of the newly developed 2A-based ratiometric polyprotein probes of membrane traffic in conjunction with semiautomated image analysis software packages for quantitative analyses. The ratiometric probes and software are available from the authors.
荧光蛋白对植物膜运输的研究方式产生了巨大影响。在此，我们回顾了这些分子在该研究领域中的应用，并讨论了特定荧光蛋白衍生物在各种应用和植物物种中的优势与缺陷。我们详细讨论了对表达水平进行定量估计的必要性，以及荧光蛋白在生物合成膜运输定量分析中的潜力。本文还提供了详细的描述和方案，介绍了新开发的基于2A的膜运输比率多蛋白探针与半自动图像分析软件包结合用于定量分析的使用方法。比率探针和软件可向作者索取。
