Brusslan J A, Karlin-Neumann G A, Huang L, Tobin E M
Department of Biology, University of California, Los Angeles 90024-1606.
Plant Cell. 1993 Jun;5(6):667-77. doi: 10.1105/tpc.5.6.667.
We analyzed a mutant of Arabidopsis with a severely reduced level of cab140 RNA. This mutant, named lct for low level of cab140 transcript, was obtained during a selection for phytochrome signal transduction mutants. The selection was based on reduced expression of the tumor morphology shoots gene (tms2), an introduced counter-selectable marker under the control of the cab140 promoter. Expression of the introduced cab140::tms2 gene was also greatly reduced in lct, but surprisingly, expression of other phytochrome-regulated genes was not comparably affected. Furthermore, the lct phenotype could not be separated genetically from the T-DNA insert; thus, we suggest that this phenotype was caused by cosuppression of the introduced construct and the endogenous cab140 gene, and that the mutation causing the cosuppression was located on the T-DNA insert. In vitro nuclear transcription experiments demonstrated that the suppression was occurring at the level of transcription. We also found that the suppressed cab140 genes were not significantly more methylated than the nonsuppressed cab140 genes.
我们分析了拟南芥中cab140 RNA水平严重降低的一个突变体。这个突变体,因cab140转录本水平低而命名为lct,是在筛选光敏色素信号转导突变体的过程中获得的。筛选基于肿瘤形态芽基因(tms2)表达的降低,tms2是一个导入的可反向选择标记,受cab140启动子控制。导入的cab140::tms2基因在lct中的表达也大幅降低,但令人惊讶的是,其他光敏色素调节基因的表达并未受到类似影响。此外,lct的表型在遗传上无法与T-DNA插入区分开来;因此,我们认为这种表型是由导入构建体和内源性cab140基因的共抑制引起的,并且导致共抑制的突变位于T-DNA插入上。体外核转录实验表明抑制发生在转录水平。我们还发现,被抑制的cab140基因与未被抑制的cab140基因相比,甲基化程度并没有显著增加。