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Sodium phosphate dibasic

磷酸氢二钠

Company: Sigma-Aldrich
Catalog#: S3264
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BMV Propagation, Extraction and Purification Using Chromatographic Methods
Author:
Date:
2018-07-20
[Abstract]  Brome mosaic virus (BMV) is a well-known plant virus representing single-stranded RNA (ssRNA) positive-sense viruses. It has been widely used as a model in multiple studies concerning plant virus biology, epidemiology and the application of viral capsids in nanotechnology. Herein, we describe a method for BMV purification based on ion-exchange and size-exclusion chromatography. The presented method is of similar efficiency to previously described protocols relying on differential centrifugation and can easily be scaled up. The resulting BMV capsids are stable and monodisperse and can be used for further applications. [摘要]  雀麦花叶病毒(BMV)是众所周知的植物病毒,代表单链RNA(ssRNA)正义病毒。 它已被广泛用作植物病毒生物学,流行病学和病毒衣壳在纳米技术中的应用的多项研究中的模型。 在本文中,我们描述了基于离子交换和尺寸排阻色谱的BMV纯化方法。 所提出的方法与先前描述的依赖于差速离心的方案具有相似的效率,并且可以容易地按比例放大。 得到的BMV衣壳是稳定的并且是单分散的,并且可以用于进一步的应用。

【背景】纳米技术要克服的关键挑战之一是制定有效的和组织特异性的药物递送方法。植物病毒和病毒样颗粒(VLP)具有生物相容性和可生物降解性,不含对人类或动物健康有害的病原体,是合成药物载体的安全替代品,通常会激活免疫系统的不良反应或积聚在免疫系统中。身体到毒性水平。最后,病毒衣壳的生产相对便宜且快速(Ren et al。,2007; Arcangeli et al。,2014)。

Bromoviridae 家族的雀麦花叶病毒(BMV)是用作纳米颗粒载体的良好候选物,因为它显示出所有上述特征并且是研究最多的植物病毒之一(Figlerowicz,2000; Alejska et al。,2005; Urbanowicz et al。,2005; Wierzchoslawski et al。,2006; Kao vet al。 ...

Bacterial Microcolonies in Gel Beads for High-throughput Screening
Author:
Date:
2018-07-05
[Abstract]  High-throughput screening of a DNA library expressed in a bacterial population for identifying potentially rare members displaying a property of interest is a crucial step for success in many experiments such as directed evolution of proteins and synthetic circuits and deep mutational scanning to identify gain- or loss-of-function mutants.

Here, I describe a protocol for high-throughput screening of bacterial (E. coli) microcolonies in gel beads. Single cells are encapsulated into monodisperse water-in-oil emulsion droplets produced with a microfluidic device. The aqueous solution also contains agarose that gelates upon cooling on ice, so that solid gel beads form inside the droplets. During incubation of the emulsion, the cells grow into monoclonal microcolonies ...
[摘要]  在细菌群体中表达的DNA文库的高通量筛选用于鉴定显示感兴趣性质的潜在稀有成员是在许多实验中成功的关键步骤,例如蛋白质和合成回路的定向进化以及用于鉴定增益的深度突变扫描 - 或功能丧失的突变体。

在这里,我描述了一种用于高通量筛选凝胶珠中细菌(大肠杆菌)微菌落的方案。将单细胞包封成用微流体装置产生的单分散油包水乳液液滴。水溶液还含有琼脂糖,其在冰上冷却时凝胶化,从而在液滴内部形成固体凝胶珠。在乳液温育期间,细胞在珠内生长成单克隆微菌落。在从乳液中分离凝胶珠并通过荧光激活细胞分选(FACS)分选后,从凝胶珠中回收细菌,然后准备进行进一步的分选,诱变或分析。为了通过FACS分类,该方案需要荧光读数,例如荧光报告蛋白的表达。测量微小菌落的平均荧光信号降低了高表型细胞间变异性的影响,并且与单细胞分选相比提高了灵敏度。我们应用这种方法在ON和OFF状态下对pBAD启动子文库进行分类(Duarte et al。,2017)。

【背景】荧光激活细胞分选(FACS)具有> 10 7 事件/ h的无与伦比的筛选通量(Davies,2012)。然而,通过FACS根据其荧光分选单个细胞以筛选合成回路的文库(Schaerli和Isalan,2013)经常受到高表型细胞间变异性的阻碍。或者,可以对水凝胶珠中所含的小细胞集落(微集落)进行分类(Weaver ...

Protocol for RYMV Inoculation and Resistance Evaluation in Rice Seedlings
Author:
Date:
2018-06-05
[Abstract]  Rice yellow mottle virus (RYMV), a mechanically transmitted virus that causes serious damage to cultivated rice plants, is endemic to Africa. Varietal selection for resistance is considered to be the most effective and sustainable management strategy. Standardized resistance evaluation procedures are required for the identification and characterization of resistance sources. This paper describes a protocol for mechanical inoculation of rice seedlings with RYMV and two methods of resistance evaluation – one based on a symptom severity index and the other on virus detection through double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA). [摘要]  非洲特有的稻黄斑驳病毒(RYMV)是一种机械传播的病毒,对栽培稻株造成严重损害。 品种抗性选择被认为是最有效和可持续的管理策略。 需要标准化的阻力评估程序来识别和表征阻力源。 本文介绍了利用RYMV机械接种水稻幼苗的方案,以及两种抗性评估方法 - 一种基于症状严重性指数,另一种基于双抗体夹心酶联免疫吸附试验(DAS-ELISA)进行病毒检测。

【背景】RYMV是非洲水稻生产的主要生物限制因素(Séréet al。,2013),在非洲和马达加斯加的大多数水稻生产国都有报道。它不能通过种子传播(Konaté等人,2001; Allarangaye等人,2006),而是通过昆虫载体(特别是甲虫)和农业作业期间的接触传播Bakker,1974;Traoré等人,2006),尤其是在将苗床的苗移植到田间时。该病毒非常稳定,能够在大米和野生禾本科(Bakker,1974)宿主中以高浓度繁殖。

监测苗床RYMV发病率和品种选择是管理RYMV最有效和可持续的方式。存在两种表型的抗性 - 部分抗性,其特征在于症状出现延迟(Albar等人,1998)和高抗性,其特征在于使用DAS-ELISA不存在病毒检测(Ndjiondjop et al。,1999)。虽然部分抗性在水稻品种中广泛分布,但栽培水稻品种仅有少数品种O.水稻和 O。 glaberrima ...

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