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KCl

氯化钾

Company: Sigma-Aldrich
Catalog#: P9333
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Rice Ragged Stunt Virus Propagation and Infection on Rice Plants
Author:
Date:
2018-10-20
[Abstract]  Virus inoculation is a basic experimental procedure to evaluate the resistance of a rice variety or a transgenic material upon virus infection. We recently demonstrated that Rice Ragged Stunt Virus (RRSV), an oryzavirus that is transmitted by brown planthopper (BPH), can suppress jasmonic acid-mediated antiviral defense through the induction of microRNA319 and facilitate virus infection in rice. To verify this, we performed virus inoculation experiments on wild-type rice plants and miR319-TCP21-associated transgenic rice plants through a modified group inoculation method. Here, we presented the detailed procedure of RRSV propagation and infection process on rice plants. [摘要]  病毒接种是评估水稻品种或转基因材料对病毒感染的抗性的基本实验程序。 我们最近证明,由褐飞虱(BPH)传播的 Rice Ragged Stunt Virus >(RRSV)是一种 oryzavirus >,可以通过诱导抑制茉莉酸介导的抗病毒防御 microRNA319并促进水稻中的病毒感染。 为了验证这一点,我们通过改良的组接种方法对野生型水稻植物和miR319-TCP21相关的转基因水稻植物进行了病毒接种实验。 在这里,我们介绍了水稻植物RRSV繁殖和感染过程的详细程序。
【背景】研究病毒发病机制和筛选抗病毒水稻品种已经在克服水稻病毒病和保持粮食安全方面做了大量工作。 在该领域中,病毒接种是评估转基因材料或水稻品种的抗性的必要且可靠的方法。 水稻粗糙特技病毒>(RRSV)可以在许多亚洲国家以持续繁殖的方式通过褐飞虱传播水稻褴褛特技病(Ling et al。>,1978; Hibino,1979)。 两种经典方法,包括单苗接种和组接种,已用于进行病毒接种实验(Zhang et al。>,2013)。 通过修改传统的群体接种方法,我们提供了一种与自然感染条件非常相似的便捷方法。

Fluorescence Titrations to Determine the Binding Affinity of Cyclic Nucleotides to SthK Ion Channels
Author:
Date:
2018-10-05
[Abstract]  The cyclic-nucleotide modulated ion channel family includes cyclic nucleotide-gated (CNG) and hyperpolarization-activated and cyclic nucleotide-modulated (HCN) channels, which play essential roles in visual and olfactory signaling and the heart pacemaking activity. Functionally, these channels have been extensively characterized by electrophysiological techniques from protein heterologously expressed in Xenopus oocytes and mammalian cells. On the other hand, expression and purification of these proteins for biophysical and structural analyses in vitro is problematic and expensive and, accordingly, only limited information on the purified channels is available in the literature. Here we describe a protocol for binding studies of fluorescently labeled cyclic nucleotides to a ... [摘要]  环核苷酸调节的离子通道家族包括环核苷酸门控(CNG)和超极化激活和环核苷酸调节(HCN)通道,其在视觉和嗅觉信号传导和心脏起搏活动中起重要作用。在功能上,这些通道已经通过来自非洲爪蟾卵母细胞和哺乳动物细胞中异源表达的蛋白质的电生理学技术进行了广泛的表征。另一方面,这些蛋白质的表达和纯化用于体外生物物理和结构分析是有问题且昂贵的,因此,文献中仅提供关于纯化通道的有限信息。在这里,我们描述了用于将荧光标记的环核苷酸与真核CNG通道的同源物结合研究的方案。此外,我们描述了如何在竞争测定中直接探测未标记的环核苷酸的结合。使用荧光作为配体结合的灵敏探针可减少所需蛋白质的量,并使用标准实验室设备进行快速简便的测量。
【背景】了解蛋白质在分子细节中的功能需要广泛的微观表征。对于配体门控离子通道,需要进行不同的分析以获得有关蛋白质与配体的特异性相互作用,配体结合位点和孔隙之间的通信以及通道特异性特征(如离子通量和失活或脱敏)的信息。属性。与对应于各种功能状态的通道构象的结构数据一起,这允许开发通道功能和调节的完整机械描述。环核苷酸门控(CNG)离子通道是四聚体钾通道,由于它们在嗅觉和视觉信号级联中的功能而特别受关注(Kaupp和Seifert,2002; Craven和Zagotta,2006)。然而,在确定的条件下,纯化的CNG通道的数据非常有限,主要来自单分子力谱(Higgins ...

In vivo and in vitro 31P-NMR Study of the Phosphate Transport and Polyphosphate Metabolism in Hebeloma cylindrosporum in Response to Plant Roots Signals
Author:
Date:
2018-08-20
[Abstract]  We used in vivo and in vitro phosphorus-31 nuclear magnetic resonance (31P-NMR) spectroscopy to follow the change in transport, compartmentation and metabolism of phosphate in the ectomycorrhizal fungus Hebeloma cylindrosporum in response to root signals originating from host (Pinus pinaster) or non-host (Zea mays) plants. A device was developed for the in vivo studies allowing the circulation of a continuously oxygenated mineral solution in an NMR tube containing the mycelia. The in vitro studies were performed on fungal material after several consecutive treatment steps (freezing in liquid nitrogen; crushing with perchloric acid; elimination of perchloric acid; freeze-drying; dissolution in an appropriate liquid ... [摘要]  我们使用体内和体外磷-31核磁共振( 31 P-NMR)光谱来跟踪运输,分区和 外生菌根真菌 Hebeloma cylindrosporum 中的磷酸盐代谢响应来自宿主( Pinus pinaster )或非宿主( Zea mays )的根信号植物。 开发了一种用于体内研究的装置,其允许连续氧化的矿物质溶液在含有菌丝体的NMR管中循环。 在几个连续的处理步骤(在液氮中冷冻;用高氯酸压碎;消除高氯酸;冷冻干燥;在适当的液体培养基中溶解)后,对真菌材料进行体外研究。

【背景】 菌根真菌和植物之间的关联改善了宿主植物的P营养(Smith和Read,2008; Plassard和Dell,2010; Cairney,2011; Smith 等人,,2015)。这种积极效应主要归因于真菌菌丝对磷酸盐(Pi)的吸收,探测了在活跃吸收根周围的耗竭区以外的大量土壤(Smith和Read,2008; Cairney,2011; Smith et al。< em="">,2015)和真菌细胞分泌细胞外磷酸酶(Quiquampoix和Mousain,2005)。吸收的Pi部分地掺入磷酸化的代谢物,磷脂和核酸中,并且部分地浓缩成多磷酸盐(PolyP),其中它们构成液泡中的储存池(Ashford 等人,,1994)。该协议详述了一种装置,该装置允许通过 31 ...

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