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Phenylmethanesulfonyl fluoride

苯基甲磺酰氟

Company: Sigma-Aldrich
Catalog#: P7626
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RNA Immunoprecipitation (RIP) Sequencing of Pri-miRNAs Associated with the Dicing Complex in Arabidopsis
Author:
Date:
2018-07-05
[Abstract]  RNA immunoprecipitation (RIP) is an antibody-based technique used to map in vivo RNA-protein interactions. DBR1, an RNA debranching enzyme, is responsible for the debranching of lariat RNA, for the degradation and turnover of lariat RNAs. It is well known that primary miRNA (Pri-miRNA) is recognized and further processed into mature miRNA by the Dicing complex mainly composed of DCL1 and HYL1. Due to the low abundance of pri-miRNAs, RIP followed qRT-PCR has been widely used to evaluate the binding efficiency of the Dicing complex with pri-miRNAs in previous studies. Therefore, the genome-wide evaluation of the Dicing complex with pri-miRNAs is lacking. With the improvement of high-throughput sequencing technologies, we successfully used RIP-seq to compare the binding efficiency ... [摘要]  RNA免疫沉淀(RIP)是一种基于抗体的技术,用于绘制体内 RNA-蛋白质相互作用。 DBR1是一种RNA脱支酶,负责套索RNA的脱支,用于套索RNA的降解和转换。众所周知,主要miRNA(Pri-miRNA)被主要由DCL1和HYL1组成的切割复合物识别并进一步加工成成熟miRNA。由于pri-miRNA的丰度较低,RIP随后的qRT-PCR已被广泛用于评估切割复合物与pri-miRNA在先前研究中的结合效率。因此,缺乏对具有pri-miRNA的切割复合物的全基因组评估。随着高通量测序技术的改进,我们成功地使用RIP-seq比较了Dicing复合物与野生型和 dbr1-2 突变体之间的pri-miRNA的结合效率。 。在该方案中,我们提供了在两种不同基因型之间的HYL1-YFP和DCL1-YFP转基因植物中使用GFP捕获珠的RIP-seq的详细描述。该方法可用于评估pri-miRNA与拟南芥中的切割复合物的结合,并且它可以应用于植物中的其他RNA结合蛋白。

Implementation of Blue Light Switchable Bacterial Adhesion for Design of Biofilms
Author:
Date:
2018-06-20
[Abstract]  Control of bacterial adhesions to a substrate with high precision in space and time is important to form a well-defined biofilm. Here, we present a method to engineer bacteria such that they adhere specifically to substrates under blue light through the photoswitchable proteins nMag and pMag. This provides exquisite spatiotemporal remote control over these interactions. The engineered bacteria express pMag protein on the surface so that they can adhere to substrates with nMag protein immobilization under blue light, and reversibly detach in the dark. This process can be repeatedly turned on and off. In addition, the bacterial adhesion property can be adjusted by expressing different pMag proteins on the bacterial surface and altering light intensity. This protocol provides light ... [摘要]  在空间和时间上高精度地控制细菌粘附到基底对于形成明确的生物膜是重要的。 在这里,我们提出了一种方法来设计细菌,使其在蓝光下通过光可切换蛋白质nMag和pMag特异性地粘附在基底上。 这为这些交互提供了精妙的时空遥控。 工程菌在表面上表达pMag蛋白,以便它们可以在蓝光下与nMag蛋白固定化的基质粘附,并在黑暗中可逆地分离。 该过程可以重复开启和关闭。 此外,通过在细菌表面表达不同的pMag蛋白质并改变光强度可以调节细菌粘附性质。 该协议提供了可高度空间和时间分辨率的细菌粘附的光可切换,可逆和可调控制,这使我们能够以极大的灵活性在基底上图案化细菌。

【背景】控制生物膜形成对于了解细菌在自然发生的生物膜中的社会相互作用至关重要(Flemming et。,2016)。这对生物膜在生物催化,生物传感和废物处理中的生物技术应用也特别重要(Zhou等人,2013; Jensen等人,2016)。生物膜的形成始终始于细菌与底物的粘附,这决定了生物膜中的空间组织(Liu等人,2016; Nadell等人,2016)。已经提出了许多策略来控制细菌粘附,例如通过脂质体融合利用生物正交反应基团修饰细菌表面(Elahipanah等,2016),将粘附分子固定在基质上(Sankaran等,等),2015; Zhang等人,2016; ...

Enzymatic Activity Assay for Invertase in Synechocystis Cells
Author:
Date:
2018-05-20
[Abstract]  Invertase can catalyze the hydrolysis of sucrose, and is widely distributed in cells of cyanobacteria and plants. Being responsible for the first step for sucrose metabolism, invertase plays important physiological roles and its enzymatic activity is frequently needed to be determined. All the methods for determination of the invertase activity are dependent on detection of the glucose product generated by the invertase. Here we describe an ion chromatography based protocol of our laboratory for determination of cyanobacterial intracellular invertase activity. [摘要]  转化酶可催化蔗糖的水解,广泛分布于蓝细菌和植物细胞中。 负责蔗糖代谢的第一步,转化酶起着重要的生理作用,其酶活性经常需要确定。 所有测定转化酶活性的方法都依赖于转化酶产生的葡萄糖产物的检测。 在这里我们描述了我们的实验室用于测定蓝细菌细胞内转化酶活性的基于离子色谱的方案。

【背景】转化酶和蔗糖在蓝细菌中发挥重要的生理作用(Curatti,et al。,2008; Kolman等人,2015)和高等植物(Vargas等人, ,2003; Vargas et。,2010)。转化酶(EC 3.2.1.26)可以催化蔗糖降解成葡萄糖和果糖。由于转化酶的这种特性,任何可用于测定葡萄糖或果糖的方法理论上都将用于转化酶酶活性测定。实际上,大多数转化酶酶活性测定基于检测产生的葡萄糖产物。

一些公司已经开发了用于转化酶活性测定的几种试剂盒,例如来自abcam(USA)的ab197005,来自Novus Biologicals(USA)的KA1629,来自Sigma-Aldrich(USA)的MAK118。通过使用这些试剂盒,由转化酶反应产生的葡萄糖产物将被氧化并通过比色(570nm)或荧光法(λem/ ex = 585 / ...

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