Activation tagging in aspen using an inducible two component Ac/Ds-enhancer element system

使用诱导型双组分 Ac/Ds 增强子元件系统在白杨中进行激活标记

• 批准号: 5449207
• 负责人: Privatdozent Dr. Matthias Fladung
• 金额: --
• 依托单位: Institut für Forstgenetik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2005
• 资助国家: 德国
• 起止时间: 2004-12-31 至 2010-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5449207?language=en
• 关键词: Activation; tagging; aspen; using; inducible; Activation; tagging; aspen; using; inducible

Based on the Ac/Ds two element transposition system from maize an activation tagging approach is suggested for the hybrid aspen (Populus tremula x tremuloides) line ¿Esch5¿. The proposed approach is based on results obtained from our earlier work on the genetic transfer of the maize transposable element Ac and its functional analysis in hybrid and pure aspen lines. It was shown that the Ac element is active in aspen and reintegrates elsewhere in genomic regions in high frequency. However, a two element transposon tagging system where Ac and Ds are put together in crosses is not feasible in trees due to the in part long vegetative phases. To overcome this barrier, an inducible two element Ac/ATDs element system is suggested to induce activation tagged variants following two independent transformation steps. In combination with a 35S enhancer tetramer and outward facing two CaMV 35S promoter located near both ends of the ATDs element, expression of genes can be elevated which are located adjacent to the new integration site of the element. As selective marker for ATDs transposition, both knocking-out the expression of a phenotypic marker (rolC gene) and a negative selection marker gene (tms) are considered. Thus, the transposition can easily be screened in primary transgenic lines.

基于玉米的AC/DS两个元素换位系统，建议用于混合aspen（populus tremula x tremuloides）线�ESCH5¿的激活标记方法。所提出的方法基于我们早期关于玉米转座元素AC遗传转移的工作及其在混合和纯白杨线中的功能分析的结果。结果表明，AC元素在ASPEN中活跃，并以高频为基因组区域的其他地方重新融合。但是，由于部分长蔬菜相，在树上将AC和DS放在十字架中的两个元素转座标记系统是不可行的。为了克服这一障碍，建议诱导的两个元素AC/ATD元素系统在两个独立的转换步骤之后诱导激活标记的变体。结合35S增强子四聚体和外在面向ATDS元素两端附近的两个CAMV 35s启动子，可以将基因的表达升高，这些表达与元素的新积分位点相邻。作为ATDS换位的选择性标记，都考虑了表型标记（ROLC基因）的表达和负选择标记基因（TMS）的表达。那就可以轻松地在主要转基因线上筛选换位。