中国科技核心期刊
CN:31-1600/Q
ISSN:1004-0374
“健康与疾病的免疫”国际学术研讨会通知      关于有网站冒充本刊网站的声明
《生命科学》 2017, 29(5): 462-467
病毒重组细菌人工染色体构建、突变及应用
刘立涛,孙洪磊,刘金华*
(中国农业大学动物医学院,北京 100193)
摘 要:如何快速、方便地保存和修饰基因组较大的DNA 和RNA 病毒核酸一直是病毒学研究的重点。细菌人工染色体(bacterial artificial chromosome, BAC) 为解决这两个问题提供了便利,它可以容纳300 kb 的基因序列,能够通过大肠杆菌诱变技术进行有效修饰,目前已经成为编辑大基因组病毒基因的有效工具。DNA 病毒,如疱疹病毒和痘病毒的基因组已被克隆到BAC 载体上。此外,一些RNA 病毒,如冠状病毒和黄病毒科病毒等也建立了基于BAC 的反向遗传操作系统。现概述不同病毒重组BAC 构建策略,总结了常用的对BACs 进行基因修饰的技术,介绍了BAC 载体重组病毒的拯救以及将BAC 序列从病毒基因组中删除的方法,简述了病毒重组细菌人工染色体的应用。
 
Viral bacterial artificial chromosomes: construction, mutagenesis and application
LIU Li-Tao, SUN Hong-Lei, LIU Jin-Hua*
(College of Veterinary Medicine, China Agriculture University, Beijing 100193, China)
Abstract: Maintenance and manipulation of large DNA and RNA virus genomes have been a focus for virological research. Bacterial artificial chromosome (BAC) vectors provids a solution as they can harbor large DNA sequences and can efficiently be modified using well-established mutagenesis techniques in Escherichia coli, and has become an effective tool for editing large genomes. Numerous DNA virus genomes of herpes virus and pox virus were cloned into BAC vectors. In addition, several reverse genetic systems for RNA viruses such as members of Coronaviridae and Flaviviridae could be established based on BAC constructs. In this paper, we provide an overview on the strategies used for the generation of virus BAC vectors and also on systems currently available for various virus species. Furthermore, we address common mutagenesis techniques that allow modification of BACs and review the reconstitution of viruses from BAC vectors and strategies of removing the bacterial sequences from the virus genome. Finally, we briefly discuss the application of viral bacterial artificial chromosomes.
 
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