用于通路(Gateway)克隆技术的植物表达载体研究进展

Research Progress on Plant Expression Vectors for Gateway Cloning Technology

通路(Gateway)克隆技术是根据λ噬菌体基因组和大肠杆菌基因组之间的位点专一性重组分子机制开发的一套分子克隆新技术。利用该技术LR反应构建目的基因的表达载体时不需要经过酶切和连接等繁琐而又费时的过程,因此,可以节省很多时间。为了扩大Gateway技术在植物基因工程领域的应用,最近有很多研究机构和研究小组开发了能用于组成型或诱导型表达目的基因、基因沉默、启动子分析、蛋白质亚细胞定位、蛋白质/蛋白质相互作用、多个DNA片段的模块化组装和DNA组片段功能验证等研究用的植物表达载体。该文对这些技术的研究进展进行了综述。

Gateway molecular cloning technology was developed by Invitrogen based on the mechanism of site-specific recombination between λ phage DNA and the Escherichia coli ge- nome. Construction of an expression vector for a target gene via Gateway LR reaction does not need to use restriction enzymes and a ligase, and thus is time efficient. Recently, to expand the application of Gateway technology in plant genetic engineering, several research groups have constructed Gateway destination vectors for studies on constitutive, inducible or silence expression of target genes, promoter analyses, protein localization, protein/protein interaction, DNA fragment modular assembly and functional verification of genomic fragments. Here we re- view research progress on the development of these destination vectors.