A New Barley Stripe Mosaic Virus Allows Large Protein Overexpression for Rapid Function Analysis
A New Barley Stripe Mosaic Virus Allows Large Protein Overexpression for Rapid Function Analysis
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DOI：
10.1104/pp.17.01412
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发表时间：
2018-03-01
期刊：
影响因子：
7.4
通讯作者：
Houde, Mario
中科院分区：
文献类型：
--
作者：
Cheuk, Arnaud;Houde, Mario
了解基因功能的遗传和分子基础对于利用它们的潜力生产具有新性状的植物越来越重要。一个重要的目标是提高植物生产力，以满足粮食作物生产的未来需求。在转基因植物中，基因功能主要通过过表达或沉默来表征。这种方法是一个漫长的过程，特别是在谷物中。植物病毒表达系统可用于蛋白质的快速表达。然而，目前的系统具有小的货物容量，并且主要用于基因沉默。在这里，一个四组分大麦条纹花叶病毒为基础的系统，具有高货物容量的重组蛋白在不同的植物物种的快速和稳定的表达，允许在不同的发展阶段的功能分析。荧光标记蛋白在1周内以高水平表达，并且使用苹果酸铝转运蛋白1基因显示了有效功能分析的证据。除了基因共转化的能力，这项工作表明，四组分大麦条纹花叶病毒为基础的系统允许过表达的cDNA高达2,100个核苷酸（编码类似的蛋白质78 kD），从而提供了一个宝贵的工具，以加快功能基因组学和蛋白质组学研究在单子叶植物和双子叶植物物种。
Understanding the genetic and molecular bases of gene function is of increasing importance to harness their potential to produce plants with novel traits. One important objective is the improvement of plant productivity to meet future demands in food crop production. Gene function is mostly characterized through overexpression or silencing in transgenic plants. This approach is a lengthy procedure, especially in cereals. Plant viral expression systems can be used for rapid expression of proteins. However, current systems have a small cargo capacity and have mostly been used for gene silencing. Here, a four-component barley stripe mosaic virus-based system with high cargo capacity was constructed for the rapid and stable expression of recombinant proteins in different plant species, allowing function analyses at different stages of development. Fluorescent marker proteins are expressed at high levels within 1 week, and a proof of efficient function analysis is shown using the aluminum malate transporter1 gene. In addition to the ability of gene cotransformation, this work demonstrates that the four-component barley stripe mosaic virus-based system allows the overexpression of cDNAs of up to 2,100 nucleotides (encoding a protein of similar to 78 kD), thereby providing an invaluable tool to accelerate functional genomics and proteomic research in monocot and dicot species.