Development of pGEMINI, a Plant Gateway Destination Vector Allowing the Simultaneous Integration of Two cDNA via a Single LR-Clonase Reaction.
Development of pGEMINI, a Plant Gateway Destination Vector Allowing the Simultaneous Integration of Two cDNA via a Single LR-Clonase Reaction.
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DOI：
10.3390/plants6040055
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发表时间：
2017-11-12
期刊：
影响因子：
--
通讯作者：
Simkin AJ
中科院分区：
文献类型：
--
作者：
Exposito-Rodriguez M;Laissue PP;López-Calcagno PE;Mullineaux PM;Raines CA;Simkin AJ
Gateway技术已用于促进产生大量用于研究目的的植物修饰的构建体。然而，许多目前可用的载体仅允许将单个目的cDNA整合到表达克隆中。组合过表达多个基因的能力对于植物发育的研究是必不可少的，其中几个转录物在一个或多个代谢过程中发挥作用。进行此类研究的工具是有限的，并且在许多情况下依赖于通过常规克隆将cDNA掺入表达系统，这可能既耗时又费力。据我们所知，这项研究报告的第一个发展的载体，允许同时整合两个独立的cDNA通过一个单一的LR-克隆酶反应。该载体“pGEMINI”代表了一种强大的分子工具，其提供了研究多cDNA构建体对植物发育的作用的能力，并通过瞬时或稳定转化程序开启了基因堆叠过程和基因组合的研究。
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 “pGEMINI” 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.
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