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古菌病毒基因组编辑和内源性病毒蛋白纯化。

Genome editing in archaeal viruses and endogenous viral protein purification.

机构信息

Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, 2200 Copenhagen N, Denmark.

出版信息

STAR Protoc. 2021 Sep 17;2(4):100791. doi: 10.1016/j.xpro.2021.100791. eCollection 2021 Dec 17.

Abstract

Archaea-infecting viruses are morphologically and genomically among the most diverse entities. Unfortunately, they are also fairly understudied due to a lack of efficient genetic tools. Here, we present a detailed protocol for the CRISPR/Cas-based genome editing of the virus SIRV2 infecting the genus , which could easily be adapted to other archaeal viruses. This protocol also includes the procedure for endogenous viral protein purification and identification, allowing for assessing the molecular mechanisms behind virus life cycle and virus-host interactions. For complete details on the use and execution of this protocol, please refer to Mayo-Muñoz et al. (2018) and Bhoobalan-Chitty et al. (2019).

摘要

感染古菌的病毒在形态和基因组上是最具多样性的实体之一。不幸的是,由于缺乏有效的遗传工具,它们的研究也相当不足。在这里,我们提供了一种基于 CRISPR/Cas 的详细方案,用于编辑感染属的病毒 SIRV2 的基因组,该方案很容易适应其他古菌病毒。该方案还包括内源性病毒蛋白的纯化和鉴定程序,可用于评估病毒生命周期和病毒-宿主相互作用背后的分子机制。有关该方案使用和执行的完整详细信息,请参见 Mayo-Muñoz 等人(2018 年)和 Bhoobalan-Chitty 等人(2019 年)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cfd/8456065/b3e72e101022/fx1.jpg

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