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Author:
Date:
2016-02-05
[Abstract] Lipid-Protein interaction assay is a method to search lipids, which are bound with proteins in vitro. Since membranes that are spotted with chloroplast lipids such as monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), and sulfoquinovosyldiacylglycerol (SQDG) are not commercially available, we extracted these lipids from cyanobacterial cells and spotted them onto membranes. The prepared membranes could be used for lipid-protein interaction assay.
[摘要] 脂质 - 蛋白质相互作用测定法是一种搜索在体外与蛋白质结合的脂质的方法。 由于用叶绿体脂质例如单半乳糖基二酰基甘油（MGDG），二半乳糖基二酰基甘油（DGDG）和磺基奎诺糖基二酰基甘油（SQDG）点样的膜不可商购，因此我们从蓝藻细胞中提取这些脂质并将它们点样到膜上。 所制备的膜可用于脂质 - 蛋白质相互作用测定。
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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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