A Set of Maize Transgenic Lines for Localized Mutagenesis Based on the Ac-Ds Transposon System Mutagenesis Based on the Ac-Ds Transposon System

一组用于基于 Ac-Ds 转座子系统的局部诱变的玉米转基因系 基于 Ac-Ds 转座子系统的诱变

• 批准号: 0211547
• 负责人: Hugo Dooner
• 金额: $ 14.06万
• 依托单位: Rutgers University New Brunswick
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-15 至 2004-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0211547&HistoricalAwards=false
• 关键词: Set; Maize; Transgenic; Lines; Localized

The goals of this NSF Plant Genome project are to demonstrate that the transposon Ac (Activator) can be used as a gene identification and isolation tool, as well as a mutagen, in the complex maize genome, and to develop the necessary tools to facilitate that use. The first goal was met in the initial phase of this project where Ac was shown to insert exclusively in or close to genes and to be, therefore, an excellent gene-searching engine in the highly repetitive maize genome. A highly embryogenic bz wx inbred line was developed and transformed with Ac* and Ds* constructs that had been modified to facilitate the PCR isolation of the transposon-adjacent sequence. The main goal of the second phase of this project is to create a comprehensive set of transgenic lines that will serve as starting points for the production of future insertion libraries. This phase will occur in two well-defined stages. The first will be to demonstrate the germinal transposition of an engineered Ac* or Ds* element and the ready isolation of DNA adjacent to the transposon. The second will be to identify a method of maize transformation that will enable the production of a useful set of transposon lines for localized mutagenesis. Once the above two objectives have been met, the last stage of the project to produce a set of 124 transgenic lines with a uniquely marked element at evenly spaced locations in the genome will be initiated. In that set, most genes in the maize genome will be within 7 cM of a launching platform and will be, therefore, realistic targets in a localized transposon mutagenesis experiment. These lines will be deposited in the Maize Stock Center and will serve as starting materials for the generation of future insertion libraries by interested scientists.

NSF植物基因组计划的目标是证明转座子Ac（激活子）可以作为复杂玉米基因组中的基因鉴定和分离工具，以及诱变剂，并开发必要的工具来促进这种使用。第一个目标在这个项目的初始阶段就实现了，Ac被证明只插入或靠近基因，因此在高度重复的玉米基因组中是一个优秀的基因搜索引擎。建立了高胚胎性的bz wx自交系，并将其转化为Ac*和Ds*结构，使其易于PCR分离转座子邻近序列。该项目的第二阶段的主要目标是创建一套全面的转基因系，作为未来插入文库生产的起点。这一阶段将分为两个明确定义的阶段。第一个将是证明一个工程的Ac*或Ds*元素的生发转位，以及转座子附近DNA的现成分离。第二个将是确定一种玉米转化方法，使其能够生产一组有用的转座子系用于局部诱变。一旦上述两个目标得到满足，项目的最后阶段将开始生产一组124个转基因系，这些转基因系在基因组中间隔均匀的位置上有一个独特的标记元素。在该集合中，玉米基因组中的大多数基因将在发射平台7厘米范围内，因此将成为局部转座子诱变实验的现实目标。这些细胞系将被储存在玉米库存中心，并将作为未来有兴趣的科学家生成插入库的起始材料。