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利用玉米Ac元件对拟南芥第三条染色体进行区域插入诱变。

Regional insertional mutagenesis on chromosome III of Arabidopsis thaliana using the maize Ac element.

作者信息

Dubois P, Cutler S, Belzile F J

机构信息

Université Laval, Québec, Canada.

出版信息

Plant J. 1998 Jan;13(1):141-51. doi: 10.1046/j.1365-313x.1998.00006.x.

Abstract

The authors describe the production and characterization of a collection of Arabidopsis lines each carrying a transposed Ac (trAc) element. A total of 507 lines were obtained following germinal transpositions of a single Ac element located on the upper portion of chromosome III. Southern analysis revealed that up to 90% of the lines in this collection harbour distinct insertions of Ac in the Arabidopsis genome. As previous studies on the behaviour of Ac in Arabidopsis have indicated that approximately two out of three transposition events occur to linked loci, the authors hypothesized that this collection could be of great use in isolating insertional mutants for genes located in the vicinity of the donor locus. PCR and phenotypic screens were performed to identify mutations in five loci located within a 40 cm region of chromosome III centered on the donor locus. Molecular analyses confirmed the presence of germinal insertions of Ac in three of the loci (NPTII, ABI3 and EST #210A22). At a fourth locus (AtDMC1), despite the absence of a germinal insertion, one line in which somatic insertions occurred regularly was identified and may be of use in isolating a germinal insertion. This collection of trAc lines constitutes a useful complement to the existing collection of T-DNA insertion lines and will soon be made available through the Arabidopsis Biological Resource Center.

摘要

作者描述了一系列拟南芥株系的产生及特性,每个株系都携带一个转座的Ac(trAc)元件。在位于第三条染色体上部的单个Ac元件发生生殖转座后,共获得了507个株系。Southern分析表明,该群体中高达90%的株系在拟南芥基因组中含有不同的Ac插入。由于先前对Ac在拟南芥中行为的研究表明,大约三分之二的转座事件发生在连锁位点,作者推测该群体在分离位于供体位点附近基因的插入突变体方面可能非常有用。进行了PCR和表型筛选,以鉴定位于以供体位点为中心的第三条染色体40厘米区域内的五个位点的突变。分子分析证实,在其中三个位点(NPTII、ABI3和EST #210A22)存在Ac的生殖插入。在第四个位点(AtDMC1),尽管没有生殖插入,但鉴定出一个体细胞插入经常发生的株系,可能有助于分离出生殖插入。这个trAc株系群体是现有T-DNA插入株系群体的有用补充,很快将通过拟南芥生物资源中心提供。

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