Earl P L, Moss B
National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, USA.
Curr Protoc Protein Sci. 2001 May;Chapter 5:Unit5.14. doi: 10.1002/0471140864.ps0514s13.
After a recombinant vaccinia virus is made, its DNA and protein products can be analyzed in several ways. Protocols are provided in this unit for identification of the recombinant virus by PCR (with verification of correct insertion of the DNA by Southern blotting) and by dot-blot hybridization. Also, when antibodies are available, protein expression can be analyzed by immunological methods detailed here such as dot blotting with an antibody, immunoblotting and/or immunoprecipitation. In addition, immunostaining can be used for identification of recombinant plaques as well as for determination of the purity of a recombinant virus stock. All of the protocols in this unit can be used for characterization of modified vaccinia virus Ankara (MVA) recombinant viruses.
制备重组痘苗病毒后，可通过多种方法对其DNA和蛋白质产物进行分析。本单元提供了通过PCR（通过Southern印迹验证DNA的正确插入）和斑点印迹杂交鉴定重组病毒的方案。此外，当有抗体时，可通过此处详述的免疫方法分析蛋白质表达，如用抗体进行斑点印迹、免疫印迹和/或免疫沉淀。另外，免疫染色可用于鉴定重组噬斑以及确定重组病毒储备液的纯度。本单元中的所有方案均可用于改良安卡拉痘苗病毒（MVA）重组病毒的鉴定。
