Rapid expression of transgenes driven by seed-specific constructs in leaf tissue: DHA production.
Rapid expression of transgenes driven by seed-specific constructs in leaf tissue: DHA production.
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DOI：
10.1186/1746-4811-6-8
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发表时间：
2010-03-11
期刊：
影响因子：
5.1
通讯作者：
Singh SP
中科院分区：
文献类型：
--
作者：
Petrie JR;Shrestha P;Liu Q;Mansour MP;Wood CC;Zhou XR;Nichols PD;Green AG;Singh SP
种子生物合成途径的代谢工程以多样化和改善作物产品质量是一个高度活跃的研究领域。由种子特异性启动子驱动的基因的验证是耗时的，因为转化的植物必须生长至成熟才能分析基因功能。在这项研究中，我们证明了种子特异性启动子驱动的基因包含在复杂的构建体可以瞬时表达的烟草叶测定系统通过共浸润拟南芥叶COTYLEDON 2（LEC 2）基因。一个真实世界的案例研究中，我们首先组装了一个有效的转基因DHA合成途径，使用传统的N。本氏花椰菜花叶病毒（CaMV）35 S驱动的叶测定，然后使用LEC 2-延伸测定来快速验证在拟南芥中稳定转化之前含有相同基因的复杂种子特异性构建体。LEC 2-扩展的N. Benthamiana测定允许叶组织中种子特异性启动子的瞬时激活。在这项研究中，我们已经使用的测定作为一个复杂的种子特异性转基因构建体的稳定转化前的快速初步筛选，一个功能，将变得越来越有用的基因工程从操作的单个基因的工程复杂的途径。我们建议，该试验将证明是有用的其他应用程序，其中快速表达的种子特异性结构驱动的转基因叶组织中寻求。
Metabolic engineering of seed biosynthetic pathways to diversify and improve crop product quality is a highly active research area. The validation of genes driven by seed-specific promoters is time-consuming since the transformed plants must be grown to maturity before the gene function can be analysed. In this study we demonstrate that genes driven by seed-specific promoters contained within complex constructs can be transiently-expressed in the Nicotiana benthamiana leaf-assay system by co-infiltrating the Arabidopsis thaliana LEAFY COTYLEDON2 (LEC2) gene. A real-world case study is described in which we first assembled an efficient transgenic DHA synthesis pathway using a traditional N. benthamiana Cauliflower Mosaic Virus (CaMV) 35S-driven leaf assay before using the LEC2-extended assay to rapidly validate a complex seed-specific construct containing the same genes before stable transformation in Arabidopsis. The LEC2-extended N. benthamiana assay allows the transient activation of seed-specific promoters in leaf tissue. In this study we have used the assay as a rapid preliminary screen of a complex seed-specific transgenic construct prior to stable transformation, a feature that will become increasingly useful as genetic engineering moves from the manipulation of single genes to the engineering of complex pathways. We propose that the assay will prove useful for other applications wherein rapid expression of transgenes driven by seed-specific constructs in leaf tissue are sought.
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DOI：
10.1073/pnas.0712364105
发表时间：
2008-02-26
影响因子：
11.1
作者：
Stone, Sandra L.;Braybrook, Siobhan A.;Harada, John J.
通讯作者：
Harada, John J.
影响因子：
1.9
作者：
Meyer, BJ;Mann, NJ;Howe, PRC
通讯作者：
Howe, PRC
影响因子：
7.2
作者：
Clough, SJ;Bent, AF
通讯作者：
Bent, AF
影响因子：
3
作者：
Robert, SS;Singh, SP;Green, AG
通讯作者：
Green, AG
影响因子：
3.8
作者：
Zhou, Xue-Rong;Robert, Stanley S.;Singh, Surinder P.
通讯作者：
Singh, Surinder P.