SGER: Gene Targeting of the Arabidopsis thaliana RBCS1B Locus

SGER：拟南芥 RBCS1B 基因座的基因靶向

• 批准号: 9727044
• 负责人: Wilhelm Gruissem
• 金额: $ 4.6万
• 依托单位: University of California-Berkeley
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 1998-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9727044&HistoricalAwards=false
• 关键词: SGER; Gene; Targeting; Arabidopsis; thaliana

9724044 Gruissem The goal of this work is to develop a tractable protocol for gene targeting in higher plants through homologous DNA recombination. Although homologous recombination transgenes has been reported in the past, the frequency has been low. The work will use a novel procedure that has been developed by this laboratory, targeting one the RbcS gene family in Arabidopsis. A recombinant form of RbcS is first introduced randomly into the genome and this substrate is then used for recombination events during early development. The result of recombination between the introduced gene and a resident gene of the RbcS gene family can be detected by the activation of a transgenic reporter. Recombination events will be screened for, via reporter gene expression, at the seedling stage. Once this system has been developed, it will provide a tool for reverse genetics that has been extremely successful in micro-organisms in the past, while it has been missing in higher plants.

9724044 Gruissem 这项工作的目标是开发一种易于处理的方案，通过同源 DNA 重组在高等植物中进行基因靶向。 尽管同源重组转基因过去已有报道，但频率较低。 这项工作将使用该实验室开发的一种新程序，针对拟南芥中的 RbcS 基因家族。 首先将 RbcS 的重组形式随机引入基因组中，然后将该底物用于早期发育过程中的重组事件。 引入的基因与RbcS基因家族的常驻基因之间的重组结果可以通过转基因报告基因的激活来检测。 将在幼苗阶段通过报告基因表达来筛选重组事件。 一旦这个系统被开发出来，它将提供一种反向遗传学工具，这种工具过去在微生物中非常成功，但在高等植物中却一直缺失。