Efficient linking and transfer of multiple genes by a multigene assembly and transformation vector system
Efficient linking and transfer of multiple genes by a multigene assembly and transformation vector system
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DOI：
10.1073/pnas.0931425100
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发表时间：
2003-05-13
影响因子：
11.1
通讯作者：
Li, BJ
中科院分区：
文献类型：
--
作者：
Lin, L;Liu, YG;Li, BJ
基础和应用研究经常需要引入和表达多个转基因。然而，目前，由于现有方法的技术限制，多基因转化非常困难。在这里，我们描述了一个载体系统，有效的多基因组装和转化。该系统由一个基于可转化人工染色体（TAC）的受体载体和两个供体载体组成。通过利用Cre/IoxP重组系统和归巢核酸内切酶，交替使用供体载体进行多轮基因组装循环，并将多个基因依次递送到TAC载体中。利用这个系统，我们创建了包含多达10个外源DNA片段的构建体。通过农杆菌介导法将6个抗病基因堆叠在一个载体中，转入水稻基因组。该系统扩展了分子遗传学研究和生物技术的努力，使多个基因的同时操作的剧目。
Introduction and expression of multiple transgenes is frequently required for basic and applied studies. However, at present, multigene transformation is very difficult due to technical limitations of existing methods. Here,we describe a vector system for efficient multigene assembly and transformation. The system consists of a transformation-competent artificial chromosome (TAC)-based acceptor vector together with two donor vectors. By exploiting the Cre/IoxP recombination system and homing endonucleases, multiple rounds of gene assembly cycling were carried out with alternate use of the donor vectors, and multiple genes were sequentially delivered into the TAC vector. With this system, we created constructs containing as many as 10 foreign DNA fragments. Multiple genes, including six resistant genes stacked in a construct, were transferred into rice genome by Agrobacterium-mediated transformation. This system extends the repertoire of molecular genetic studies and biotechnological endeavors by enabling simultaneous manipulation of multiple genes.