SGER: Gene Targeting of the Arabidopsis thaliana RBCS1B Locus

本工作的目标是通过同源DNA重组为高等植物的基因靶向开发一种可操作的方案。虽然同源重组转基因在过去也有报道，但频率很低。这项工作将使用该实验室开发的一种新方法，针对拟南芥中的一个红细胞基因家族。首先将重组形式的红细胞随机引入基因组，然后在早期发育期间将这种底物用于重组事件。通过激活一个转基因报告基因，可以检测导入基因与红细胞基因家族中的一个常驻基因之间的重组结果。重组事件将在苗期通过报告基因表达进行筛选。一旦这个系统被开发出来，它将为过去在微生物中非常成功的反向遗传学提供一个工具，而它在高等植物中却缺失了。

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.