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cOmplete His-Tag purification resin

Company: Roche Diagnostics
Catalog#: 5893682001
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Self-organization Assay for Min Proteins of Escherichia coli in Micro-droplets Covered with Lipids
Author:
Date:
2020-03-20
[Abstract]  The Min system determines the cell division plane of bacteria. As a cue of spatiotemporal regulation, the Min system uses wave propagation of MinD protein (Min wave). Therefore, the reconstitution of the Min wave in cell-sized closed space will lead to the creation of artificial cells capable of cell division. The Min waves emerge via coupling between the reactions among MinD, MinE, and ATP and the differences in diffusion rate on the cell membrane and in the cytoplasm. Because Min waves appear only under the balanced condition of the reaction-diffusion coupling, special attentions are needed towards several technical points for the reconstitution of Min waves in artificial cells. This protocol describes a technical method for stably generating Min waves in artificial cells. [摘要]  [摘要 ] Min系统确定细菌的细胞分裂平面。作为时空调节的提示,Min系统使用MinD 蛋白的波传播(Min wave)。因此,Min波在细胞大小的封闭空间中的重构将导致能够分裂细胞的人造细胞的产生。闵波出现经由耦合之间反应小号中MinD的,的MinE ,和ATP 和所述differenc ES 在细胞膜上的扩散速度和在细胞质中。因为最小波仅在反应扩散耦合的平衡条件下出现, 特别关注,需要对几个技术要点为闽波在人造细胞重建。该协议描述了一种在人造细胞中稳定产生Min波的技术方法。

[背景 ] 敏系统,它决定了细胞分ER 对称细胞分裂,是在细菌细胞内的组织系统的最显着的例子之一(Rothfield 等人,2005;和罗利特马戈林,2013年)。敏系统使用图案形成在细胞内的时间依赖性蛋白梯度的公知的作为敏波(宽松等人,2008; Halatek和Frey,2012;邦尼等人,2013; Zieske 。等人,2016 ; Kohyama 。等人, 2019 )。Min波是由两种蛋白MinD 和MinE 的反应扩散耦合产生的。通过与ATP结合,MinD 形成二聚体并附着在膜上。的MinE 被招募到的ATP MinD的和诱导ATP酶的活性MinD的。通过MinE ,ATP- MinD 变为ADP- MinD ,并从膜上脱离。ADP- MinD的被转换回ATP- ...

In vitro Analysis of Ubiquitin-like Protein Modification in Archaea
Author:
Date:
2018-05-20
[Abstract]  The ubiquitin-like (Ubl) protein is widely distributed in Archaea and involved in many cellular pathways. A well-established method to reconstitute archaeal Ubl protein conjugation in vitro is important to better understand the process of archaeal Ubl protein modification. This protocol describes the in vitro reconstitution of Ubl protein modification and following analysis of this modification in Haloferax volcanii, a halophilic archaeon serving as the model organism. [摘要]  泛素样(Ubl)蛋白广泛分布于古细菌中并参与许多细胞途径。 为了更好地理解古细菌Ub1蛋白质修饰的过程,重建体外古细菌Ubl蛋白质缀合物的完善方法是很重要的。 该协议描述了Ubl蛋白质修饰的体外重建以及在作为模型生物的嗜盐古细菌Haloferax volcanii 中对这种修饰进行分析。

【背景】泛素(Ub)与靶蛋白共价连接的过程被称为泛素化,其控制真核细胞中大量的细胞过程(Glickman和Ciechanover,2002; Komander和Rape,2012)。遍在蛋白化由一系列酶(包括Ub激活酶(E1),Ub结合酶(E2s)和Ub连接酶(E3s))催化。泛素化的体外重建是确定酶之间或E3与蛋白质底物之间特异性的有用测定法(Zhao等人,2012)。在古细菌中,Ubl蛋白SAMP采用Ub折叠,并且与E1样酶UbaA催化的蛋白靶标异肽连接[Maupin-Furlow,(2014)综述]。尽管E1同系物在古细菌中广泛存在,但基于一级序列比较,在大多数古细菌中未预测经典E2或E3酶。我们最近对Haloferax volcanii的研究表明甲硫氨酸亚砜还原酶A(MsrA)是Ubl蛋白质修饰(sampylation)与UbaA一起在体内温和的氧化条件下和< (体外)(fu="">

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