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The maize transposable element system Ac/Ds as a mutagen in Arabidopsis: identification of an albino mutation induced by Ds insertion.

机译:玉米转座因子系统Ac / Ds作为拟南芥中的诱变剂:鉴定由Ds插入引起的白化突变。

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摘要

A two-component transposon system based on the Ac element of maize was used as a mutagen in Arabidopsis thaliana. Transposition of a Ds element marked with a hygromycin-resistance gene was activated from four different locations in the Arabidopsis genome. The progeny of 201 plants carrying independent transposition events were screened for mutants with severe, visible phenotypes. Seven mutants were identified and four of them were analyzed genetically. Three mutations were shown to be very closely linked to a transposed copy of the element. Moreover, a mutation (alb3) causing an albino phenotype was conclusively shown to be caused by insertion of the Ds element: somatic and germinal reversion of the mutation occurred in the presence of the transposase gene but not in its absence, and in three revertants the Ds had excised from its position in the mutant line. The DNA adjacent to Ds in the mutant was isolated and it was demonstrated that revertants retained part of the 8-bp duplication caused by insertion of Ds. These experiments indicate that the Ac/Ds system can be used as an insertional mutagen in the heterologous host Arabidopsis, which will permit the isolation of genes from this species by transposon tagging.
机译:基于玉米中Ac元素的两成分转座子系统被用作拟南芥中的诱变剂。从拟南芥基因组中的四个不同位置激活了标记有潮霉素抗性基因的Ds元素的转座。筛选具有独立转座事件的201株植物的后代,以筛选具有严重可见表型的突变体。确定了七个突变体,并对其中四个进行了遗传分析。结果表明,三个突变与该元素的转座拷贝紧密相关。此外,结论性地表明,导致白化病表型的突变(alb3)是由Ds元件的插入引起的:突变的体细胞和生发回复在转座酶基因存在的情况下发生,但在不存在转座酶基因的情况下发生,在三个回复子中Ds已从其在突变株系中的位置切除。分离了突变体中与Ds相邻的DNA,并证明了回复子保留了由于插入Ds而引起的8bp重复部分。这些实验表明,Ac / Ds系统可用作异源宿主拟南芥中的插入诱变剂,这将允许通过转座子标记从该物种中分离基因。

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