Earl P L, Moss B, Wyatt L S, Carroll M W
National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, USA.
Curr Protoc Mol Biol. 2001 May;Chapter 16:Unit16.17. doi: 10.1002/0471142727.mb1617s43.
This unit first describes how to infect cells with vaccinia virus and then transfect them with a plasmid-transfer vector to generate a recombinant virus. Methods are also presented for purifying vaccinia virus and for isolating viral DNA, which can be used during transfection. Also presented are selection and screening methods used to isolate recombinant viruses and a method for the amplification of recombinant viruses. Finally, a method for live immunostaining that has been used primarily for detection of recombinant modified vaccinia virus Ankara (MVA) is presented.
本单元首先描述如何用痘苗病毒感染细胞，然后用质粒转移载体转染细胞以产生重组病毒。还介绍了纯化痘苗病毒和分离病毒DNA的方法，这些方法可用于转染过程。还介绍了用于分离重组病毒的筛选方法以及重组病毒的扩增方法。最后，介绍了一种主要用于检测重组改良安卡拉痘苗病毒（MVA）的活细胞免疫染色方法。
