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

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Ac/Ds双元件转座系统，提出了白杨<$5 <$$>的激活标记方法。所提出的方法是基于从我们早期的玉米转座因子Ac的遗传转移及其在杂交和纯白杨系中的功能分析的工作中获得的结果。结果表明，Ac元件在白杨中是有活性的，并且以高频率重新整合到基因组区域的其他地方。然而，其中Ac和Ds在杂交中放在一起的两个元件转座子标记系统在树木中是不可行的，这是由于部分长的营养生长期。为了克服这一障碍，一个可诱导的两个元件Ac/ATDs元件系统，建议诱导激活标记的变体后，两个独立的转化步骤。与位于ATDs元件两端附近的35 S增强子四聚体和面向外的两个CaMV 35 S启动子组合，可以提高位于该元件的新整合位点附近的基因的表达。作为ATDs转座的选择标记，考虑敲除表型标记（rolC基因）和阴性选择标记基因（tms）的表达。因此，可以容易地在初级转基因系中筛选转座。