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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Double in situ Hybridization
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Author:
Date:
2012-02-05
[Abstract] Double in situ hybridization I very useful to examine the relationship between the expression of two genes. But it is tricky because of the cross reaction of two different antibodies. This protocol is a valid method to do double color in situ hybridization in zebrafish embryos.
[摘要] 双重原位杂交我非常有用于检查两个基因的表达之间的关系。 但它是棘手的,因为两个不同的抗体的交叉反应。 此协议是在斑马鱼胚胎中进行双重染色原位杂交的有效方法。
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In situ Hybridization in Zebrafish Embryos
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Author:
Date:
2012-02-05
[Abstract] In situ hybridization is routinely used to examine the gene expression level and location of embryos. This protocol is modified from the Thisse protocol and is a detailed description of the in situ hybridization procedures in zebrafish embryos.
[摘要] 原位杂交常规用于检查胚胎的基因表达水平和位置。 该协议是从Thisse协议修改的,并且是斑马鱼胚胎中原位杂交程序的详细描述。
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