Activation tagging using the En-I maize transposon system in Arabidopsis
Activation tagging using the En-I maize transposon system in Arabidopsis
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DOI：
10.1104/pp.003327
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发表时间：
2002-08-01
期刊：
影响因子：
7.4
通讯作者：
Pereira, A
中科院分区：
文献类型：
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作者：
Marsch-Martinez, N;Greco, R;Pereira, A
利用玉米（Zen mays） En-I转座子系统，建立了一种在拟南芥中产生稳定激活标签插入的方法。该方法采用温室选择性标记基因，可有效地产生大量插入。大约8300个独立稳定的激活标签插入已经产生。在温室中对一组包含约2900个插入的植物进行突变筛选，发现约31个显性突变，表明显性突变频率约为1%。从第一批在温室中筛选的约400个稳定插入物中，鉴定出4个功能增益、显性激活标记、形态突变体。描述了一种名为thread的新功能获得突变体，其靶基因与通过T-DNA激活标记鉴定的YUCCA黄素单加氧酶属于同一家族。在群体中发现的功能增益突变的高频率表明，这里描述的En-I系统是一种有效的策略，可以用激活标签插入来使植物基因组饱和。由于只需要少量的初级转化体来产生激活标签种群，因此En-I系统似乎是研究目前转化方法效率较低的植物物种的一个有吸引力的替代方案。
A method for the generation of stable activation tag inserts was developed in Arabidopsis using the maize (Zen mays) En-I transposon system. The method employs greenhouse selectable marker genes that are useful to efficiently generate large populations of insertions. A population of about 8,300 independent stable activation tag inserts has been produced. Greenhouse-based screens for mutants in a group of plants containing about 2,900 insertions revealed about 31 dominant mutants, suggesting a dominant mutant frequency of about 1%. From the first batch of about 400 stable insertions screened in the greenhouse, four gain-in-function, dominant activation-tagged, morphological mutants were identified. A novel gain-in-function mutant called thread is described, in which the target gene belongs to the same family as the YUCCA flavin-mono-oxygenase that was identified by T-DNA activation tagging. The high frequency of identified gain-in-function mutants in the population suggests that the En-I system described here is an efficient strategy to saturate plant genomes with activation tag inserts. Because only a small number of primary transformants are required to generate an activation tag population, the En-I system appears to be an attractive alternative to study plant species where the present transformation methods have low efficiencies.