构建作为细菌人工染色体的冠状病毒感染性cDNA
Engineering infectious cDNAs of coronavirus as bacterial artificial chromosomes.
作者信息
Almazán Fernando, Márquez-Jurado Silvia, Nogales Aitor, Enjuanes Luis
机构信息
Department of Molecular and Cell Biology, Centro Nacional de Biotecnología (CNB-CSIC), Campus Universidad Autónoma de Madrid, Darwin 3, Cantoblanco, Madrid, 28049, Spain.
出版信息
Methods Mol Biol. 2015;1282:135-52. doi: 10.1007/978-1-4939-2438-7_13.
Abstract
The large size of the coronavirus (CoV) genome (around 30 kb) and the instability in bacteria of plasmids carrying CoV replicase sequences represent serious restrictions for the development of CoV infectious clones using reverse genetic systems similar to those used for smaller positive sense RNA viruses. To overcome these problems, several approaches have been established in the last 13 years. Here we describe the engineering of CoV full-length cDNA clones as bacterial artificial chromosomes (BACs), using the Middle East respiratory syndrome CoV (MERS-CoV) as a model.
摘要
冠状病毒（CoV）基因组较大（约30 kb），且携带CoV复制酶序列的质粒在细菌中不稳定，这严重限制了使用类似于用于较小正链RNA病毒的反向遗传系统来开发CoV感染性克隆。为克服这些问题，在过去13年里已建立了几种方法。在此，我们以中东呼吸综合征冠状病毒（MERS-CoV）为模型，描述了将CoV全长cDNA克隆构建为细菌人工染色体（BAC）的工程方法。
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