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Magnesium Sulfate Heptahydrate (MgSO4·7H2O)

硫酸镁七水合物

Company: Sigma-Aldrich
Catalog#: 63140
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Expression, Purification and Crystallization of the Herpesvirus Nuclear Egress Complex (NEC)
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Date:
2016-07-20
[Abstract]  The protocol describes the production and crystallization of the soluble form of the nuclear egress complex (NEC) from Herpes simplex virus 1 and Pseudorabies virus. The NEC is a heterodimer that consists of conserved proteins UL31 and UL34. NEC oligomerization deforms the inner nuclear membrane around the capsid in infected cells, thereby mediating capsid budding into the perinuclear space during nuclear egress. We have successfully developed a protocol for large-scale preparation of highly pure NEC from two different viruses in a prokaryotic expression system, which enabled us to crystallize these viral protein complexes and determine their structures. This procedure may be adapted to purify and crystallize other soluble protein complexes. [摘要]  该协议描述了来自单纯疱疹病毒1和伪狂犬病病毒的核出口复合物(NEC)的可溶形式的产生和结晶。 NEC是由保守蛋白UL31和UL34组成的异源二聚体。 NEC低聚使受感染细胞中的衣壳周围的内核膜变形,从而在核出口期间介导衣壳发芽进入核周空间。 我们已经成功地开发了一个协议,从大规模制备高纯度NEC从两种不同的病毒在原核表达系统,这使我们能够结晶这些病毒蛋白复合物和确定其结构。 该程序可适于纯化和结晶其它可溶性蛋白质复合物。

Identification and Characterization of Bacterial Chemoreceptors Using Quantitative Capillary and Gradient Plate Chemotaxis Assays
Author:
Date:
2016-04-20
[Abstract]  Bacterial chemotaxis is a motility-based response that biases cell movement toward beneficial molecules, called attractants, and away from harmful molecules, also known as repellents. Since the species of the genus Pseudomonas are characterized by a metabolic versatility, these bacteria have developed chemotactic behaviors towards a wide range of different compounds. The specificity of a chemotactic response is determined by the chemoreceptor, which is at the beginning of the signaling cascade and which receives the signal input. The basic elements of a typical chemoreceptor are the periplasmic ligand binding domain (LBD), responsible for sensing environmental stimuli, and the cytosolic methyl-accepting (MA) domain, that interacts with other components of the cellular signaling cascade. Escherichia ... [摘要]  细菌趋化性是基于运动性的反应,其将细胞运动偏向有益分子,称为引诱剂,并远离有害分子,也称为驱避剂。由于假单胞菌属的种属的特征在于代谢多样性,这些细菌已经对宽范围的不同化合物产生趋化行为。趋化反应的特异性由化学感受器确定,化学感受器处于信号级联的开始并且接受信号输入。典型的化学感受器的基本元件是负责感测环境刺激的周质配体结合结构域(LBD)和与细胞信号传导级联的其它组分相互作用的细胞溶质甲基接受(MA)结构域。大肠杆菌(大肠杆菌)是趋化性研究中的传统模型,具有5个良好表征的化学感受器。然而,基因组序列分析已经揭示许多其他细菌具有更多的化学感受器,其中一些具有部分重叠的信号谱。这种高数目的化学感受器通过分析单个化学感受器突变体使他们的研究变得复杂。我们已经寻求化学受体表征的替代策略,其对应于由所研究的化学受体的LBD和E的MA结构域组成的嵌合受体的产生。大肠杆菌受体(Tar)。然后将嵌合体引入E的无化学感受器的突变体。并且所得菌株的趋化性完全是由于该嵌合受体的作用。在本公开中,我们描述了使用定量毛细管和梯度板测定来研究假单胞菌趋化性以及E。含有嵌合受体的大肠杆菌菌株。

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