Department of Biological Sciences, University of South Carolina, Columbia, SC, USA.
Department of Biology and Geology, University of South Carolina Aiken, Aiken, SC, USA.
Methods Mol Biol. 2023;2686:351-363. doi: 10.1007/978-1-0716-3299-4_18.
The β-glucuronidase (GUS) reporter gene system is an important technique with versatile uses in the study of flower development in a broad range of species. Transcriptional and translational GUS fusions are used to characterize gene and protein expression patterns, respectively, during reproductive development. Additionally, GUS reporters can be used to map cis-regulatory elements within promoter sequences and to investigate whether genes are regulated post-transcriptionally. Gene trap/enhancer trap GUS constructs can be used to identify novel genes involved in flower development and marker lines useful in mutant characterization. Flower development studies primarily have used the histochemical assay in which inflorescence tissue from transgenic plants containing GUS reporter genes are stained for GUS activity and examined as whole-mounts or subsequently embedded into wax and examined as tissue sections. In addition, quantitative GUS activity assays can be performed on either floral extracts or intact flowers using a fluorogenic GUS substrate. Another use of GUS reporters is as a screenable marker for plant transformation. A simplified histochemical GUS assay can be used to quickly identify transgenic tissues.
β-葡糖苷酸酶（GUS）报告基因系统是一种在广泛物种的花发育研究中具有多种用途的重要技术。转录和翻译 GUS 融合分别用于分别在生殖发育过程中表征基因和蛋白质表达模式。此外，GUS 报告基因可用于在启动子序列内定位顺式调控元件，并研究基因是否受到转录后调控。基因捕获/增强子捕获 GUS 构建体可用于鉴定参与花发育的新基因以及在突变体表征中有用的标记品系。花发育研究主要使用组织化学分析，其中含有 GUS 报告基因的转基因植物的花序组织被染色以检测 GUS 活性，并作为整体或随后嵌入蜡中作为组织切片进行检查。此外，可使用荧光 GUS 底物对花提取物或完整花朵进行定量 GUS 活性测定。GUS 报告基因的另一个用途是作为植物转化的可筛选标记。简化的组织化学 GUS 分析可用于快速鉴定转基因组织。
