Rojas-Pierce Marcela, Springer Patricia S
Department of Botany and Plant Sciences, Center for Plant Cell Biology, University of California-Riverside, Riverside, CA, USA.
Methods Mol Biol. 2003;236:221-40. doi: 10.1385/1-59259-413-1:221.
Gene traps and enhancer traps provide a valuable tool for gene discovery. With this system, genes can be identified based solely on the expression pattern of an inserted reporter gene. The use of a reporter gene, such as beta-glucuoronidase (GUS), provides a very sensitive assay for the identification of tissue- and cell-type specific expression patterns. In this chapter, protocols for examining and documenting GUS reporter gene activity in individual lines are described. Methods for the amplification of sequences flanking transposant insertions and subsequent molecular and genetic characterization of individual insertions are provided.
基因捕获和增强子捕获为基因发现提供了一种有价值的工具。利用这个系统，仅根据插入的报告基因的表达模式就能鉴定基因。使用报告基因，如β-葡萄糖醛酸酶（GUS），为鉴定组织和细胞类型特异性表达模式提供了一种非常灵敏的检测方法。在本章中，描述了检测和记录单个株系中GUS报告基因活性的实验方案。还提供了转座子插入侧翼序列的扩增方法以及单个插入片段的后续分子和遗传特征分析方法。
