Exposito-Rodriguez Marino, Laissue Philippe P, López-Calcagno Patricia E, Mullineaux Philip M, Raines Christine A, Simkin Andrew J
School of Biological Sciences, Wivenhoe Park, University of Essex, Colchester CO4 3SQ, UK.
Genetics, Genomics and Breeding, NIAB EMR, New Road, East Malling, Kent ME19 6BJ, UK.
Plants (Basel). 2017 Nov 12;6(4):55. doi: 10.3390/plants6040055.
Gateway technology has been used to facilitate the generation of a large number of constructs for the modification of plants for research purposes. However, many of the currently available vectors only allow the integration of a single cDNA of interest into an expression clone. The ability to over-express multiple genes in combination is essential for the study of plant development where several transcripts have a role to play in one or more metabolic processes. The tools to carry out such studies are limited, and in many cases rely on the incorporation of cDNA into expression systems via conventional cloning, which can be both time consuming and laborious. To our knowledge, this study reports on the first development of a vector allowing the simultaneous integration of two independent cDNAs via a single LR-clonase reaction. This vector "" represents a powerful molecular tool offering the ability to study the role of multi-cDNA constructs on plant development, and opens up the process of gene stacking and the study of gene combinations through transient or stable transformation procedures.
网关技术已被用于促进大量构建体的生成,以便出于研究目的对植物进行修饰。然而,目前许多可用载体仅允许将单个感兴趣的cDNA整合到表达克隆中。在植物发育研究中,组合过表达多个基因的能力至关重要,因为在一个或多个代谢过程中有多个转录本发挥作用。开展此类研究的工具有限,而且在许多情况下依赖于通过传统克隆将cDNA掺入表达系统,这既耗时又费力。据我们所知,本研究首次报道了一种载体的开发,该载体允许通过单个LR克隆酶反应同时整合两个独立的cDNA。该载体“”是一种强大的分子工具,能够研究多cDNA构建体在植物发育中的作用,并通过瞬时或稳定转化程序开启基因叠加和基因组合研究的进程。