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

拟南芥研究团体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，基础设施将不会完成：（1）研究人员必须能够获得基因组中每个基因的零突变体。 (2)必须创造出能够敲除串联重复基因的工具。 为了满足这些需求，将开发由含有Ds转座子发射台和Cre/Lox重组位点的映射T-DNA系的集合组成的反向遗传资源。 这些功能元件将使用户能够创建基因组任何片段的靶向缺失，无论是一组串联重复的基因还是目前不存在突变等位基因的单个基因。 通过先前的努力，10，450个这些pDs-Lox T-DNA系被递送到俄亥俄州州立大学的拟南芥生物资源中心。 本项目的目标是在接下来的两年中向拟南芥群落提供另外50，000个映射的pDs-Lox T-DNA系。 关于这些生产线的生产和可用性的详细信息，可在以下网站上找到：www.biotech.wisc.edu/Arabidopsis。 这个项目将帮助拟南芥社区实现2010年项目的目标，为科学家提供获得基因和基因组的无效突变体，否则将无法获得。更广泛的影响：这个社区资源将为科学家提供他们需要的工具，在整个拟南芥基因组中产生有针对性的删除。 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，以伴随这一新的pDs-Lox图谱。 本教程将为用户提供与pDs-Lox群体的使用相关的理论和实践信息。 将制定详细的实验室方案，指导用户完成以下过程：（1）进行遗传杂交;（2）动员Ds元件并筛选重新插入;（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没有转座子经验的实验室提供他们需要从pDs-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，从而为本科生提供了一个机会，以获得宝贵的经验，作为研究团队的一部分工作。

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.