基孔肯雅病毒的反向遗传学方法

Reverse Genetics Approaches for Chikungunya Virus.

作者信息

Phuektes Patchara, Chu Justin Jang Hann

机构信息

Department of Pathobiology, Faculty of Veterinary Medicine, Khon Kaen University, Khon Kaen, Thailand.

Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.

出版信息

Methods Mol Biol. 2016;1426:283-95. doi: 10.1007/978-1-4939-3618-2_26.

DOI:10.1007/978-1-4939-3618-2_26

PMID:27233281

Abstract

Reverse genetic systems based on an infectious cDNA clone, a double-stranded copy of the viral genome carried on a plasmid vector, have greatly enhanced the understanding of RNA virus biology by facilitating genetic manipulation of viral RNA genomes. To date, infectious cDNA clones of Chikungunya virus (CHIKV) have been constructed using different combinations of plasmid vectors and/or bacterial host strains. Here, we describe our approaches for the construction of infectious cDNA clones of CHIKV and the protocol for genetic manipulation of the clones by site-directed mutagenesis.

摘要

基于感染性 cDNA 克隆的反向遗传系统，即携带在质粒载体上的病毒基因组双链拷贝，通过促进对病毒 RNA 基因组的遗传操作，极大地增进了对 RNA 病毒生物学的理解。迄今为止，已使用质粒载体和/或细菌宿主菌株的不同组合构建了基孔肯雅病毒（CHIKV）的感染性 cDNA 克隆。在此，我们描述了构建 CHIKV 感染性 cDNA 克隆的方法以及通过定点诱变对克隆进行遗传操作的方案。