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首页> 外文期刊>Reviews in medical virology >Cloning of herpesviral genomes as bacterial artificial chromosomes.
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Cloning of herpesviral genomes as bacterial artificial chromosomes.

机译：疱疹病毒基因组的克隆为细菌人工染色体。

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摘要

Herpesviruses, which are important pathogens for both animals and humans, have large and complex genomes with a coding capacity for up to 225 open reading frames (ORFs). Due to the large genome size and the slow replication kinetics in vitro of some herpesviruses, mutagenesis of viral genes in the context of the viral genome by conventional recombination methods in cell culture has been difficult. Given that mutagenesis of viral genes is the basic strategy to investigate function, many of the herpesvirus ORFs could not be defined functionally. Recently, a completely new approach for the construction of herpesvirus mutants has been developed, based on cloning of the virus genome as a bacterial artificial chromosome (BAC) in E. coli. This technique allows the maintenance of viral genomes as a plasmid in E. coli and the reconstitution of viral progeny by transfection of the BAC plasmid into eukaryotic cells. Any genetic modification of the viral genome in E. coli using prokaryotic recombination proteins is possible, thereby allowing the generation of mutant viruses and facilitating the analysis of herpesvirus genomes cloned as infectious BACs. In this review, we describe the principle of cloning a viral genome as a BAC using murine gammaherpesvirus 68 (MHV-68), a mouse model for gammaherpesvirus infections, as an example.
机译：疱疹病毒是动物和人类的重要病原体，具有庞大而复杂的基因组，其编码能力高达225个开放阅读框（ORF）。由于一些疱疹病毒的大基因组大小和体外慢的复制动力学，在细胞培养中通过常规重组方法诱变病毒基因组背景下的病毒基因非常困难。鉴于病毒基因的诱变是研究功能的基本策略，因此许多疱疹病毒ORF不能在功能上进行定义。最近，基于在大肠杆菌中克隆病毒基因组作为细菌人工染色体（BAC），已经开发出一种用于构建疱疹病毒突变体的全新方法。通过将BAC质粒转染到真核细胞中，该技术可以使病毒基因组作为大肠杆菌中的质粒得以维持，并可以重组病毒后代。使用原核重组蛋白对大肠杆菌中的病毒基因组进行任何遗传修饰都是可能的，从而可以产生突变病毒，并有助于分析克隆为感染性BAC的疱疹病毒基因组。在这篇综述中，我们描述了使用鼠γ疱疹病毒68（MHV-68）（一种用于γ疱疹病毒感染的小鼠模型）克隆病毒基因组作为BAC的原理。

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