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  • 毕惠惠.单子叶植物RNA干扰和过表达Gateway载体的构建[J].植物遗传资源学报,2013,14(1):117-125.    [点击复制]
  • bi huihui.Construction of RNAi and Overexpression Vectors for Genetic Transformation of Monocotyledon Plants Based on Gateway Technology[J].植物遗传资源学报,2013,14(1):117-125.   [点击复制]
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单子叶植物RNA干扰和过表达Gateway载体的构建
毕惠惠
0
(西北农林科技大学农学院)
摘要:
基因枪和农杆菌介导的遗传转化是目前常用的两种单子叶植物遗传转化方法。载体的发展和改良是提高植物遗传转化效率的重要基础, RNA干扰载体和过表达载体是目前用于植物基因功能研究的主要工具。Gateway克隆技术是一种基于lambda噬菌体特异位点重组特性的通用克隆技术,该技术可以将大批的目的基因方便、快捷地连接到受体载体上。本文利用Gateway技术结合传统的酶切、连接方法,构建了适用于单子叶植物基因枪和农杆菌转化的RNA干扰Gateway载体pAHC-PSK-RNAi、pClean-G185-RNAi和过表达Gateway载体pAHC-PSK-OE和pClean-G185-OE,为利用基因枪和农杆菌介导的遗传转化,在小麦和水稻等单子叶植物中进行规模化基因功能研究奠定了基础。
关键词:  单子叶植物  遗传转化  RNA干扰  过表达  Gateway
DOI:
投稿时间:2012-01-18修订日期:2012-04-09
基金项目:转基因专项项目2011ZX08010-003
Construction of RNAi and Overexpression Vectors for Genetic Transformation of Monocotyledon Plants Based on Gateway Technology
Abstract:
Particle bombardment and Agrobacterium-mediated transformation are the two most widely employed methods in plant genetic transformation. Vector system plays an important role in development of an efficient transgenic technology. At present, RNAi and overexpression vectors are widely used for gene function analysis. Gateway is a kind of easy and quick cloning technology, which is based on site-specific recombination characteristics of lambda phage. A number of genes can be cloned into destination vectors quickly and conveniently by using this technique. In this study, the overexpression vectors pAHC-PSK-OE and pClean-G185-OE, and the RNAi vectors pAHC-PSK-RNAi and pClean-G185-RNAi for biolistic and Agrobacterium-mediated transformation were constructed based on traditional enzyme digestion and ligation method, and gateway technology, respectively. This laid the basis for high-throughput platform for gene function analysis in monocotyledon plants.
Key words:  Monocotyledon plants  Genetic transformation  RNAi  Overexpression  Gateway

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