A Two Component Activator/Dissociation Platform for Reverse and Forward Genetic Analysis in Maize

基因组测序计划已经为理解单子叶植物(水稻)和双子叶植物(拟南芥)物种提供了蓝图，但只有不到10%的已测序植物基因的功能被实验确定。玉米是美国最重要的经济作物，也是世界范围内的主要农作物。对玉米基因功能的了解对于制定一个全面的框架以提高这一全球粮食、饲料和能源的生产率至关重要。转座子激活子(Ac)和解离(DS)将被用来创建一个平台，用于根据植物表型或DNA序列在遗传筛选中识别基因敲除。使用经典的基因技术和非转基因材料，我们将产生一个由10,000个家庭组成的集合，每个家庭都有一个均匀分布在整个基因组中的唯一的D插入。DS元件两侧的DNA序列将被克隆和测序，为玉米基因组中的每一次插入提供准确的物理位置。由于D倾向于移动到基因组中紧密相连的位置，每个D将作为针对关联基因的额外几轮突变的平台。获得项目结果该项目将与正在进行的玉米基因组测序项目和通过爱荷华州立大学和博伊斯·汤普森研究所的两个项目网站产生的数据进行整合。所有产生的基因材料将在项目期间由Brutnell实验室分发，并在项目结束时在玉米遗传合作社库存中心免费提供。

Genome sequencing projects have provided blueprints to understanding both monocot (rice) and dicot (Arabidopsis) species, yet the function of less than 10% of sequenced plant genes has been experimentally determined. Maize is the economically most important plant in the US and a major crop world-wide. An understanding of maize gene function is essential to develop a comprehensive framework for increasing the productivity of this global food, feed and energy resource. The transposable elements Activator (Ac) and Dissociation (Ds) will be utilized to create a platform for identifying gene knockouts in genetic screens based on plant phenotype or DNA sequence. Using classical genetic techniques and non-transgenic materials we will generate a collection of 10,000 families that each harbor a unique Ds insertion distributed evenly throughout the genome. DNA Sequences flanking the Ds elements will be cloned and sequenced providing a precise physical location for each insertion in the maize genome. As Ds tends to move to closely linked site sites in the genome, each Ds will serve as a platform for additional rounds of mutagenesis targeting linked genes.Access to project outcomesThis project will be integrated with ongoing maize genome sequencing projects and the data disseminated as it is generated through two project websites at Iowa State University and the Boyce Thompson Institute. All genetic materials generated will be distributed by the Brutnell lab during the project period and made freely available at the Maize Genetics Cooperative Stock Center at the end of the project period.