| The Application of Quercetin to Study the Effect of Hsp70 Silencing on Plant Virus Infection in Nicotiana benthamiana Plants
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Author:
Date:
2015-12-05
[Abstract] Pepino mosaic virus (PepMV) is a mechanically-transmitted pathogen affecting tomato plants worldwide. Like with other plant viruses (Verchot, 2012), the heat shock cognate protein 70 homolog (Hsc70) was identified as an interactor of the PepMV coat protein (CP) (Mathioudakis et al., 2012). Here, we describe a pharmacological approach to silence Hsp70 in plants using quercetin (Mathioudakis et al., 2014), an Hsp70 protein expression flavonoid inhibitor (Hosokawa et al., 1990; Manwell and Heikkila 2007). In the case of Hsp70, this methodology represents a faster and easier approach than silencing of Hsp70 by reverse genetics assays, such as VIGS methodology. Fully expanded leaves of 2 to 3 weeks old Nicotiana benthamiana plants were ...
[摘要] Pepino花叶病毒（ PepMV ）是一种机械传播的病原体，影响全世界的番茄植株。与其他植物病毒一样（Verchot，2012），热休克同源蛋白70同源物（Hsc70）被鉴定为PepMV外壳蛋白（CP）的相互作用物（Mathioudakis等人，2012）。在这里，我们描述了使用槲皮素使沉默Hsp70在植物中的药理学方法（Mathioudakis等人，2014），Hsp70蛋白质表达类黄酮抑制剂（Hosokawa等人，1990 ; Manwell和Heikkila 2007）。在Hsp70的情况下，该方法表示比通过反向遗传学测定（例如VIGS方法）沉默Hsp70更快和更容易的方法。使用注射器用槲皮素（溶解于DMSO中）或DMSO（对照植物）浸润2至3周龄的烟草烟草的完全膨胀的叶子。用PepMV病毒接种物机械接种植物。在接种后4天通过免疫印迹分析在局部叶子上分析Hsp70和PepMV的积累。
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| Western Blotting for Staphylococcus aureus AgrA
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Author:
Date:
2014-03-20
[Abstract] Staphylococcus aureus has a quorum sensing system to regulate the expression of various virulence factors, which is exerted by the agr locus that encodes agrBDCA and a regulatory RNA called RNAIII. AgrB, AgrD, and AgrC proteins are involved in producing and recognizing extracellular quorum sensing molecules and transduce the signal by altering the phosphorylation status of AgrA, which is a positive transcription factor, to regulate cytolysin genes as well as the RNAIII gene. RNAIII regulates the expression of various virulence genes. Expression of the agr locus has been examined in depth at the transcriptional level, but investigations of translational expression are limited, because immunoglobulin G used to detect a specific protein ...
[摘要] 金黄色葡萄球菌具有调节各种毒力因子的表达的群体感应系统，其通过编码agrBDCA的agr基因座和调节agrBDCA的基因座发挥作用。 RNA称为RNAIII。 AgrB，AgrD和AgrC蛋白参与产生和识别胞外群体感应分子并通过改变作为阳性转录因子的AgrA的磷酸化状态来转导信号，以调节细胞溶素基因以及RNAIII/em>基因。 调节各种毒力基因的表达。已经在转录水平上深入研究了 agr 基因座的表达，但是翻译表达的研究是有限的，因为用于检测特异性蛋白的免疫球蛋白G对于高度反应。 aureus 蛋白A.在这里，我们报告检测AgrA的方法，所述AgrA是由 agr 调节系统编码的转录因子。虽然这是用于Western印迹的特定方案。 aureus AgrA蛋白，它也可以用于其他的。金黄色葡萄球菌通过使用适当的抗体。
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