| Construction of FWPV Chimaeric MVA
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Author:
Date:
2015-01-05
[Abstract] Construction of chimaeric MVA is a useful tool with which to study gene function of related viruses. The protocol given here describes MVA chimaeras containing genes from Fowlpox virus (FWPV), although this can be applied to DNA derived from other organisms. There are a number of steps required to make the chimaeric MVA: 1) Purification of viral particles; 2) Extraction of DNA from purified viral particles; 3) Assembly of linear recombination templates; 4) Transfection of linear recombination templates; 5) Selection of chimaeric MVA.
Note: This procedure uses live virus, and should be conducted using Good Microbiological Practice, in accordance with international and national biocontainment requirements. This procedure also involves Genetic Modification of microorganisms, and ...
[摘要] 嵌合MVA的构建是研究相关病毒的基因功能的有用工具。 本文给出的方案描述了含有来自鸡痘病毒（FWPV）的基因的MVA嵌合体，尽管这可以应用于来自其他生物体的DNA。 制备嵌合MVA需要许多步骤：1）病毒颗粒的纯化; 2）从纯化的病毒颗粒中提取DNA; 3）装配线性重组模板; 4）线性重组模板的转染; 5）嵌合MVA的选择。
注意：此程序使用活病毒，并应根据国际和国家生物承诺要求使用良好微生物实践进行。 该程序还涉及微生物的遗传修饰，并且在开始前应获得适当的安全批准。
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| Construction of Deletion-knockout Mutant Fowlpox Virus (FWPV)
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Author:
Date:
2014-05-20
[Abstract] The construction of deletion-knockout poxviruses is a useful approach to determining the function of specific virus genes. This protocol is an adaptation of the transient dominant knockout selection protocol published by Falkner and Moss (1990) for use with vaccinia virus. The protocol makes use of the dominant selectable marker Escherichia coli guanine phosphoribosyltransferase (gpt) gene (Mulligan and Berg, 1981), under the control of an early/late poxvirus promoter. The deletion viruses that are produced no longer contain a selectable marker, which may be preferable for the production of vaccines.
[摘要] 缺失敲除痘病毒的构建是确定特定病毒基因的功能的有用方法。 该方案是由Falkner和Moss（1990）公布的用于牛痘病毒的瞬时显性敲除选择方案的改编。 该方案利用在早期/晚期痘病毒启动子控制下的显性选择标记大肠杆菌鸟嘌呤磷酸核糖基转移酶（gpt）基因（Mulligan和Berg，1981）。 产生的缺失病毒不再含有可选择标记，其可优选用于疫苗的生产。
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