LB-agar

LB琼脂

公司名称: EMD Millipore

产品编号: VM459383236

Preparation and Purification of β-1,3-glucan-Linked Candida glabrata Cell Wall Proteases by Ion-Exchange Chromatography, Gel Filtration, and MDPF-Gelatin-Zymography Assay

Pirjo Pärnänen, Timo Sorsa, Taina Tervahartiala and Pirjo Nikula-Ijäs

Published online: 2024-03-20

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Purification and TEM of Afp and Its Variants

Author:

Daria Rybakova, Alok K. Mitra and Mark R. H. Hurst,

Date:

2014-05-20

[Abstract] The Serratia entomophila antifeeding prophage (Afp), forms a phage-tail-like particle (tailocin) that causes cessation of feeding activity of the New Zealand grass grub, Costelytra zealandica. Here, we describe the more detailed purification protocol for Afp particles and its variants which is based on the procedure described in our original publication (Rybakova et al., 2013). The purification procedure includes inducing Escherichia coli (E. coli) cells harbouring afp genes under arabinose-inducible promoter for 24 h. The cells are ... [摘要] 噬菌体噬菌体抗原噬菌体（Afp）形成噬菌体尾样颗粒（太古霉素），其导致新西兰草禾本科（Costelytra zealandica）的饲养活性的停止。在这里，我们描述了Afp颗粒及其变体的更详细的纯化方案，其基于我们的原始出版物中描述的程序（Rybakova等人，2013）。纯化程序包括在阿拉伯糖诱导型启动子下诱导含有emp 基因的大肠杆菌（大肠杆菌）细胞24小时。收获细胞，在冰上超声处理，然后进行DNA酶处理和离心。然后将上清液过滤灭菌，并应用于尺寸排阻色谱（SEC）柱。合并含有Afp或其变体的级分并超速离心。除去上清液，将透明沉淀物重悬浮于残余缓冲液中。该程序产生约70％纯的Afp颗粒或其变体。使用透射电子显微镜（TEM）对Afp颗粒进行负染色并可视化。

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