Cotter Catherine A, Earl Patricia L, Wyatt Linda S, Moss Bernard
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.
Curr Protoc Protein Sci. 2017 Aug 1;89:5.12.1-5.12.18. doi: 10.1002/cpps.34.
The culturing of cell lines used with vaccinia virus, both as monolayer and in suspension, is described. The preparation of chick embryo fibroblasts (CEF) is presented for use in the production of the highly attenuated and host range-restricted modified vaccinia virus Ankara (MVA) strain of vaccinia virus. Protocols for the preparation, titration, and trypsinization of vaccinia virus stocks, as well as viral DNA preparation and virus purification methods are also included. © 2017 by John Wiley & Sons, Inc.
本文描述了用于痘苗病毒的细胞系在单层和悬浮培养中的培养方法。介绍了鸡胚成纤维细胞（CEF）的制备方法，用于生产高度减毒且宿主范围受限的痘苗病毒安卡拉改良株（MVA）。还包括痘苗病毒储备液的制备、滴定和胰蛋白酶消化的方案，以及病毒DNA制备和病毒纯化方法。© 2017约翰威立父子公司。
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