Purification of Herpesvirus Virions and Capsids
疱疹病毒和病毒衣壳的纯化
发布: 2014年08月05日第4卷第15期 DOI: 10.21769/BioProtoc.1193 浏览次数: 17266
下载 PDF 
Q&A
参见作者原研究论文
The authors used this protocol in:
Aug 2013
Abstract
This protocol was designed for large-scale purification of herpesvirus particles by cell culture. Virions and capsids are isolated from extracellular culture media and cell nuclei, respectively. Purity and concentration of the purified samples are usually sufficient for structural studies with cryo electron microscopy and cryo electron tomography. The protocol should also be generally suitable for purifying herpesvirus virions and capsids for other types of studies.
Keywords: Virus purification (病毒纯化)
Herpesvirus (疱疹病毒)
Density gradient centrifuge (密度梯度离心)
Materials and Reagents
- Herpesvirus infected cells (e.g., Vero cells for herpes simplex virus culture, MRC5 cells for human cytomegalovirus culture)
- PBS (pH 7.4)
- Sucrose
- NP-40
- Dry ice
- Dulbecco’s modified eagle medium (DMEM)
- Fetal bovine serum (FBS)
- Herpesvirus culture media (see Recipes)
Equipment
- Laminar flow hood
- Centrifuge with cool function
- Ultracentrifuge
- Ultra-Clear 38.5 ml tubes (fit in SW28 rotor) (Beckman Coulter, catalog number: 344058 )
- Ultra-Clear 13.2 ml tubes (fit in SW41 rotor) (Beckman Coulter, catalog number: 344059 )
- KimwipesTM tissue paper
- Tweezers
- Gradient master (Biocomp Instruments, catalog number: 107-201M )
- Flashlight
- Vortex
- 37 °C water bath
- Syringe with 23 gauge hypodermic needle
Procedure
文章信息
版权信息
© 2014 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
- Dai, X. and Zhou, Z. H. (2014). Purification of Herpesvirus Virions and Capsids. Bio-protocol 4(15): e1193. DOI: 10.21769/BioProtoc.1193.
- Dai, X., Yu, X., Gong, H., Jiang, X., Abenes, G., Liu, H., Shivakoti, S., Britt, W. J., Zhu, H., Liu, F. and Zhou, Z. H. (2013). The smallest capsid protein mediates binding of the essential tegument protein pp150 to stabilize DNA-containing capsids in human cytomegalovirus. PLoS Pathog 9(8): e1003525.
Download Citation in RIS Format
分类
微生物学 > 微生物细胞生物学 > 细胞分离和培养
微生物学 > 微生物-宿主相互作用 > 病毒
细胞生物学 > 细胞分离和培养 > 细胞生长
您对这篇实验方法有问题吗？
在此处发布您的问题，我们将邀请本文作者来回答。同时，我们会将您的问题发布到Bio-protocol Exchange，以便寻求社区成员的帮助。
提问指南
+ 问题描述
写下详细的问题描述，包括所有有助于他人回答您问题的信息（例如实验过程、条件和相关图像等）。
发布问题
