Davies John P, Reddy Vaka S, Liu Xing L, Reddy Avutu S, Ainley William Michael, Folkerts Otto, Marri Pradeep, Jiang Ke, Wagner Douglas Ry
Dow AgroSciences Indianapolis Indiana.
Present address: Molecular Microbiology School of Medicine Washington University in St Louis St Louis Missouri.
Plant Direct. 2019 Feb 21;3(2):e00118. doi: 10.1002/pld3.118. eCollection 2019 Feb.
Activation Tagging, distributing transcriptional enhancers throughout the genome to induce transcription of nearby genes, is a powerful tool for discovering the function of genes in plants. We have developed a transposable element system to distribute a novel activation tagging element throughout the genome of maize. The transposon system is built from the Enhancer/Suppressor (/) transposon system and uses an engineered seed color marker to show when the transposon excises. Both somatic and germinal excision events can be detected by the seed color. The activation tagging element is in a -derived non-autonomous transposon and contains four copies of the ugarane acilliform irus-enhancer (-enhancer) and the AAD1 selectable marker. We have demonstrated that the transposon can give rise to germinal excision events that can re-integrate into non-linked genomic locations. The transposon has remained active for three generations and events displaying high rates of germinal excision in the T2 generation have been identified. This system can generate large numbers of activation tagged maize lines that can be screened for agriculturally relevant phenotypes.
激活标签技术,即将转录增强子分布于整个基因组以诱导附近基因转录,是发现植物基因功能的强大工具。我们开发了一种转座子系统,用于在玉米基因组中分布一种新型激活标签元件。该转座子系统基于增强子/抑制子(En/Spm)转座子系统构建,并使用一种工程化的种子颜色标记来显示转座子何时切除。种子颜色可检测体细胞和生殖细胞切除事件。激活标签元件位于一个衍生的非自主转座子中,包含四个豇豆花叶病毒增强子(CaMV-enhancer)拷贝和AAD1选择标记。我们已经证明,该转座子可引发生殖细胞切除事件,这些事件可重新整合到非连锁的基因组位置。该转座子在三代中一直保持活性,并且已鉴定出在T2代中显示出高生殖细胞切除率的事件。该系统可产生大量激活标签玉米品系,可针对农业相关表型进行筛选。