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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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