van Haaren M J, Ow D W
Department of Genetics, Biocentrum Amsterdam, Netherlands.
Plant Mol Biol. 1993 Nov;23(3):525-33. doi: 10.1007/BF00019300.
The concept of gene identification and cloning using insertional mutagenesis is well established. Many genes have been isolated using T-DNA transformation or transposable elements. Maize transposable elements have been introduced into heterologous plant species for tagging experiments. The behaviour of these elements in heterologous hosts shows many similarities with transposon behaviour in Zea mays. Site-specific recombination systems from lower organisms have also been shown to function efficiently in plant cells. Combining transposon and site-specific recombination systems in plants would create the possibility to induce chromosomal deletions. This 'transposition-deletion' system could allow the screening of large segments of the genome for interesting genes and may also permit the cloning of the DNA corresponding to the deleted material by the same site-specific recombination reaction in vitro. This methodology may provide a unique means to construct libraries of large DNA clones derived from defined parts of the genome, the phenotypic contribution of which is displayed by the mutant carrying the deletion.
利用插入诱变进行基因鉴定和克隆的概念已得到充分确立。许多基因已通过T-DNA转化或转座元件分离出来。玉米转座元件已被引入异源植物物种用于标签实验。这些元件在异源宿主中的行为与玉米中转座子的行为有许多相似之处。来自低等生物的位点特异性重组系统也已被证明在植物细胞中能有效发挥作用。在植物中结合转座子和位点特异性重组系统将有可能诱导染色体缺失。这种“转座-缺失”系统可以筛选基因组的大片段以寻找感兴趣的基因，还可能通过体外相同的位点特异性重组反应克隆与缺失物质相对应的DNA。这种方法可能提供一种独特的手段来构建源自基因组特定部分的大型DNA克隆文库，其表型贡献由携带缺失的突变体显示出来。
