Delivery of multiple transgenes to plant cells
Delivery of multiple transgenes to plant cells
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DOI：
10.1104/pp.107.106104
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发表时间：
2007-12-01
期刊：
影响因子：
7.4
通讯作者：
Tzfira, Tzvi
中科院分区：
文献类型：
--
作者：
Dafny-Yelin, Mery;Tzfira, Tzvi
利用转基因技术对植物物种进行基因操作，对基础植物研究和生物技术产生了深远的影响。例如，异源基因的过表达被广泛用于将新性状引入转基因作物。过表达也可以与下调和受控表达（例如诱导的、发育调节的或组织特异性的）研究组合使用，作为基础植物研究和各种模式植物中天然基因的功能分析的工具。外源基因在植物细胞中瞬时和/或稳定表达需要两个关键成分。第一种是质粒载体，其能够携带外源基因编码序列和调节元件，即启动子和终止子序列，其在植物细胞中表达所需（因此统称为表达盒）。第二种是生物或物理方法，通过这些方法可以将载体递送到靶植物细胞中。除了病毒介导的基因表达，其中质粒载体和载体本身之间的区别是相当模糊的，生物射弹，聚乙二醇和农杆菌介导的基因转移都需要质粒载体，其中一些已经被专门设计为通过特定方法的最佳使用。因此，例如，小的和多拷贝的质粒是用于基因枪和聚乙二醇介导的遗传转化的最有用的载体，其中在每个实验中通常使用大量的DNA。另一方面，携带某些生物学特征的单拷贝质粒已被专门设计用于农杆菌作为转化载体。在过去的几十年中，已经开发出了许多适合于在植物细胞中克隆、转移和表达外源基因的质粒（例如Simoens等人，1986; Hajdukiewicz等人，1994;汉密尔顿，1997; Xiang等人，1999; Hellens等人，2000 b;厄利等人，2006; Coutu等人，2007年）。虽然这些质粒已被证明在植物生物学研究和生物技术中是有用的，但这些载体中的许多载体的基本设计是相当限制性的，并且很少允许将多于单个靶基因（不包括选择基因，其可以存在或不存在于转化载体上）作为单个单元克隆和转移到植物细胞中。对解剖和分析复杂代谢途径的兴趣日益增长，并且需要开发植物生物技术的多基因性状的全部潜力（综述参见Halpin等人，2001; Halpin和Boerjan，2003; Capell和Christou，2004; Tyo等人，2007）授权开发用于在植物细胞中递送和堆叠多个基因的新方法和工具。在这里，我们回顾了一些方法，可用于将多个基因导入植物细胞，而主要集中在系统，允许组装的多基因植物转化载体。
The use of transgenic technologies for the genetic manipulation of plant species has had a profound impact on basic plant research and biotechnology. Overexpression of heterogonous genes, for example, is widely used for the introduction of novel traits into transgenic crop plants. Overexpression can also be used in combination with down-regulation and controlled expression (eg induced, developmentally regulated, or tissue specific) studies as a tool for basic plant research and for functional analysis of native genes in various model plants. Two crucial components are required for the transient and/or stable expression of foreign genes in plant cells. The first is a plasmid vector capable of carrying the foreign gene-encoding sequence and the regulatory elements, ie promoter and terminator sequences, needed for its expression in plant cells (thus collectively referred to as an expression cassette). The second is the biological or physical methods by which the vector can be delivered into the target plant cells. With the exception of virusmediated gene expression, where the distinction between the plasmid vector and the vehicle itself is rather blurry, biolistics, polyethylene glycol, andAgrobacteriummediated gene transfer all require plasmid vectors, some of which have been specifically designed for optimal use by a particular method. Thus, for example, small and multicopy plasmids are most useful vectors for biolistics and polyethylene glycol-mediated genetic transformation where large quantities of DNA are typically used in each experiment. On the other hand, single-copy plasmids carrying certain biological features have been specifically designed for use with Agrobacterium as the transformation vehicle. In the past several decades, a remarkable variety of plasmids suitable for the cloning, transfer, and expression of foreign genes in plant cells has been developed (eg Simoens et al., 1986; Hajdukiewicz et al., 1994; Hamilton, 1997; Xiang et al., 1999; Hellens et al., 2000b; Earley et al., 2006; Coutu et al., 2007). While these plasmids have proven instrumental in plant biology research and biotechnology, the basic designs of many of these vectors are quite restrictive and rarely permit the cloning and transfer of more than a single target gene (excluding the selection gene, which may or may not be present on the transformation vector) as a single unit into plant cells. The growing interest in dissecting and analyzing complex metabolic pathways and the need to exploit the full potential of multigene traits for plant biotechnology (for review, see Halpin et al., 2001; Halpin and Boerjan, 2003; Capell and Christou, 2004; Tyo et al., 2007) mandate the development of new methods and tools for the delivery and stacking of multiple genes in plant cells. Here, we review some of the methods that can be used for the delivery of multiple genes into plant cells, while focusing mainly on systems that allow the assembly of multigene plant transformation vectors.