Farrar Kerrie, Evans I Marta, Topping Jennifer F, Souter Martin A, Nielsen John E, Lindsey Keith
The Integrative Cell Biology Laboratory, School of Biological and Biomedical Sciences, University of Durham, South Road, Durham DH1 3LE, UK.
Plant J. 2003 Jan;33(1):61-73. doi: 10.1046/j.1365-313x.2003.01608.x.
To identify new genes expressed in meristematic cells, a promoter trap insertional mutagenesis strategy was used in Arabidopsis thaliana. Transgenic line AtEM201 exhibits promoter trap GUS activity in embryos and in the regions of active cell division in the seedling, notably the apical meristems and young leaves. The tagged gene was named EXORDIUM (EXO). AtEM201 contains a single copy of the promoter trap T-DNA, located in the EXO gene promoter, resulting in a much reduced level of EXO transcription. Seedlings homozygous for the T-DNA insertion have no obvious mutant phenotype. The EXO gene, which forms part of a small gene family in Arabidopsis, is structurally related to the tobacco PHI-1 gene, which is re-activated in cultured cells following release from phosphate starvation-induced cell cycle arrest. Expression of both the EXO-GUS and the native EXO genes is downregulated by exogenous cytokinin. Expression studies using semisynchronised cells suggest that EXO mRNA is preferentially abundant during M phase of the cell cycle. Double mutant studies revealed that the exo mutation can suppress the defective root meristem phenotype of the hydra2 mutant, suggesting that EXO may be a component of a negative regulatory system for cell division.
为了鉴定在分生细胞中表达的新基因,在拟南芥中采用了启动子捕获插入诱变策略。转基因株系AtEM201在胚胎以及幼苗中活跃细胞分裂的区域,特别是顶端分生组织和幼叶中表现出启动子捕获GUS活性。被标记的基因被命名为EXORDIUM(EXO)。AtEM201含有位于EXO基因启动子中的单个启动子捕获T-DNA拷贝,导致EXO转录水平大幅降低。T-DNA插入纯合的幼苗没有明显的突变表型。EXO基因是拟南芥中一个小基因家族的一部分,在结构上与烟草PHI-1基因相关,后者在从磷酸盐饥饿诱导的细胞周期停滞中释放后在培养细胞中重新激活。EXO-GUS和天然EXO基因的表达都受到外源细胞分裂素的下调。使用半同步细胞的表达研究表明,EXO mRNA在细胞周期的M期优先丰富。双突变体研究表明,exo突变可以抑制hydra2突变体有缺陷的根分生组织表型,这表明EXO可能是细胞分裂负调控系统的一个组成部分。