Arabidopsis 2010: A Community Resource for Generating Targeted Deletions Throughout the Arabidopsis Genome

至关重要的是，拟南芥研究界必须完成建立其反向遗传基础设施的工作。在满足两个重要标准之前，这个基础设施是不完整的：(1)研究人员必须能够获得基因组中每个基因的零突变。(2)必须创造出能够将串联复制的基因敲除的工具。为了满足这些需求，将开发一种反向遗传资源，该资源由一系列包含Ds转座子发射台和Cre/Lox重组位点的T-DNA序列组成。这些功能元件将使用户能够创建任何基因组片段的靶向删除，无论是一组串联复制的基因还是目前不存在突变等位基因的单个基因。通过之前的努力，10450个这些pDs-Lox T-DNA系被送到了俄亥俄州立大学的拟南芥生物资源中心。目前项目的目标是在未来两年内向拟南芥社区提供额外的50,000个pDs-Lox T-DNA图谱。有关这些生产线的生产和可用性的详细信息可在以下网站找到：www.biotech.wisc.edu/Arabidopsis。该项目将帮助拟南芥群落实现2010年项目的目标，为科学家提供获取基因和基因组的零突变体的途径，否则这些基因和基因组将无法获得。更广泛的影响：这个社区资源将为科学家提供他们需要在整个拟南芥基因组中产生目标缺失的工具。为了确保最广泛的研究人员能够使用这项技术，将开发一个全面的基于网络的教程，以配合这个新的pDs-Lox系列。本教程将为用户提供与pDs-Lox种群的使用相关的理论和实践信息。将制定详细的实验室协议，指导用户完成以下过程：(1)进行遗传杂交；(2)调动一个Ds元素并筛选重新插入；(3)进行Cre/Lox重组并筛选所需的缺失。这些定制方案的可用性将为以前没有转座子经验的实验室提供他们需要从pd - lox系统中受益的信息。这个基于网络的教程将由本科生开发，从而为本科生提供一个机会，获得作为研究团队一部分的宝贵经验。

It is essential that the Arabidopsis research community finish the job of building its reverse-genetic infrastructure. That infrastructure will not be complete until two important criteria are satisfied: (1) Researchers must have access to a null-mutant of every gene in the genome. (2) Tools must be created that allow tandemly-duplicated genes to be knocked-out. In order to satisfy these needs, a reverse-genetic resource consisting of a collection of mapped T-DNA lines that contain Ds transposon launch-pads and Cre/Lox recombination sites will be developed. These functional elements will give users the ability to create targeted deletions of any segment of the genome, be it a group of tandemly-duplicated genes or a single gene for which no mutant allele currently exists. Through previous efforts, 10,450 of these pDs-Lox T-DNA lines were delivered to the Arabidopsis Biological Resource Center at Ohio State University. The goal of the present project is to deliver an additional 50,000 mapped pDs-Lox T-DNA lines to the Arabidopsis community over the course of the next two years. Details regarding the production and availability of these lines can be found at the following web site: www.biotech.wisc.edu/Arabidopsis. This project will help the Arabidopsis community achieve the goals of the 2010 project by providing scientists with access to null mutants for genes and groups of genes that would otherwise not be available.Broader Impacts: This community resource will provide scientists with the tools that they need to generate targeted deletions throughout the Arabidopsis genome. In order to ensure that this technology is accessible to the broadest range of researchers, a comprehensive, web-based tutorial will be developed to accompany this new collection of mapped pDs-Lox lines. This tutorial will provide users with both theoretical and practical information relating to the uses of the pDs-Lox population. Detailed lab protocols will be developed that guide users through the following processes: (1) performing genetic crosses; (2) mobilizing a Ds element and screening for re-insertions; (3) performing Cre/Lox recombination and screening for the desired deletion. The availability of these tailor-made protocols will give labs with no prior transposon experience the information that they need to benefit from the pDs-Lox system. This web-based tutorial will be developed by undergraduate students, thereby providing an opportunity for undergraduates to gain valuable experience working as part of a research team.