Sundaresan V, Springer P, Volpe T, Haward S, Jones J D, Dean C, Ma H, Martienssen R
Cold Spring Harbor Laboratory, New York 11724, USA.
Genes Dev. 1995 Jul 15;9(14):1797-810. doi: 10.1101/gad.9.14.1797.
The crucifer Arabidopsis thaliana has been used widely as a model organism for the study of plant development. We describe here the development of an efficient insertional mutagenesis system in Arabidopsis that permits identification of genes by their patterns of expression during development. Transposable elements of the Ac/Ds system carrying the GUS reporter gene have been designed to act as enhancer traps or gene traps. A novel selection scheme maximizes recovery of unlinked transposition events. In this study 491 plants carrying independent transposon insertions were generated and screened for expression patterns. One-half of the enhancer trap insertions and one-quarter of the gene trap insertions displayed GUS expression in seedlings or flowers, including expression patterns specific to organs, tissues, cell types, or developmental stages. The patterns identify genes that act during organogenesis, pattern formation, or cell differentiation. Transposon insertion lines with specific GUS expression patterns provide valuable markers for studies of Arabidopsis development and identify new cell types or subtypes in plants. The diversity of gene expression patterns generated suggests that the identification and cloning of Arabidopsis genes expressed in any developmental process is feasible using this system.
十字花科植物拟南芥已被广泛用作研究植物发育的模式生物。我们在此描述了一种在拟南芥中高效的插入诱变系统的开发，该系统可通过基因在发育过程中的表达模式来鉴定基因。携带GUS报告基因的Ac/Ds系统转座元件已被设计用作增强子陷阱或基因陷阱。一种新颖的筛选方案可最大程度地提高非连锁转座事件的回收率。在本研究中，产生了491株携带独立转座子插入的植物，并对其表达模式进行了筛选。一半的增强子陷阱插入和四分之一的基因陷阱插入在幼苗或花中显示出GUS表达，包括器官、组织、细胞类型或发育阶段特有的表达模式。这些模式可鉴定在器官发生、模式形成或细胞分化过程中起作用的基因。具有特定GUS表达模式的转座子插入系为拟南芥发育研究提供了有价值的标记，并鉴定了植物中的新细胞类型或亚型。所产生的基因表达模式的多样性表明，使用该系统鉴定和克隆在任何发育过程中表达的拟南芥基因是可行的。
