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A rapid and efficient method for uniform gene expression using the barley stripe mosaic virus

机译:利用大麦条纹花叶病毒快速有效地进行基因均匀表达的方法

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摘要

BackgroundThe barley stripe mosaic virus (BSMV) has become a popular vector to study gene function in cereals. However, studies have been limited to gene silencing in leaves of barley or wheat. In addition, the method produces high variability between different leaves and plants. To overcome these limitations, we explored the potential of modifying the inoculation protocol for BSMV gene overexpression. An improved light, oxygen or voltage-sensing (iLOV) domain-based fluorescent protein was used as a reporter of gene expression to monitor the infection and spread of BSMV. Tobacco (Nicotiana benthamiana) leaves were infected via agroinfiltration and the leaves were homogenized to extract the BSMV particles and inoculate wheat tissues using the traditional leaf abrasion method or by incubation during seed imbibition in a Petri dish.
机译:背景大麦条纹花叶病毒(BSMV)已成为研究谷物基因功能的流行载体。但是,研究仅限于大麦或小麦叶片中的基因沉默。另外,该方法在不同的叶子和植物之间产生很大的变异性。为克服这些限制,我们探索了修改BSMV基因过表达接种方案的潜力。一种改进的基于光,氧或电压感应(iLOV)域的荧光蛋白被用作基因表达的报告基因,以监测BSMV的感染和传播。烟草(Nicotiana benthamiana)叶片通过农杆菌渗入法感染,然后将叶片均质化以提取BSMV颗粒,并使用传统的叶片磨蚀方法或在种子在Petri盘中吸水期间的孵育中接种小麦组织。

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