A Gateway-based platform for multigene plant transformation
A Gateway-based platform for multigene plant transformation
复制标题
DOI：
10.1007/s11103-006-9065-3
复制
发表时间：
2006-12-01
影响因子：
5.1
通讯作者：
Wang, Xue-Chen
中科院分区：
文献类型：
--
作者：
Chen, Qi-Jun;Zhou, Hai-Meng;Wang, Xue-Chen
后基因组时代为多基因性状、多性状和多基因产物的遗传操作提供了无与伦比的机会。然而，剩余的技术障碍使得在植物中操纵多个基因变得困难。在这里，我们描述了一个基于网关的载体系统，使多个转基因直接连接或融合。该载体系统由一个目的载体和两个特殊的attL侧翼的入口载体组成，每个入口载体含有与attL不相容的attR盒。通过多轮LR重组反应，我们称之为MultiRound Gateway，通过交替使用两种特殊的入口载体，可以将多个转基因顺序地和无限期地递送到Gateway兼容的目的载体中。在我们的原理验证实验中，我们使用该载体系统构建了一个植物转化载体，该载体包含7个功能性DNA片段，包括一个筛选标记基因、两个报告基因和四个基质附着区序列。该系统为充分发挥植物遗传操作的潜力提供了一个平台。
The post-genomic era offers unrivalled opportunities for genetic manipulation of polygenic traits, multiple traits, and multiple gene products. However, remaining technical hurdles make the manipulation of multiple genes in plants difficult. Here we describe a Gateway-based vector system to enable multiple transgenes to be directly linked or fused. The vector system consists of a destination vector and two special attL-flanked entry vectors each containing an attR cassette incompatible with the attL. By multiple rounds of LR recombination reactions, which we call MultiRound Gateway, multiple transgenes can be delivered sequentially and indefinitely into the Gateway-compatible destination vector through alternate use of the two special entry vectors. In our proof-of-principle experiments we have used this vector system to construct a plant transformation vector containing seven functional DNA fragments, including a screening marker gene, two reporter genes and four matrix attachment region sequences. This system provides a platform for fully realizing the potential of plant genetic manipulation.