Engineering Infectious cDNAs of Coronavirus as Bacterial Artificial Chromosomes
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.
Key wordsCoronavirus MERS Reverse genetics Infectious clones Bacterial artificial chromosomes
This work was supported by grants from the Ministry of Science and Innovation of Spain (MCINN) (BIO2010-16705), the European Community’s Seventh Framework Programme (FP7/2007–2013) under the project “EMPERIE” (HEALTH-F3-2009-223498), and the National Institute of Health (NIH) of the USA (2P01AI060699-06A1). S. M. received a predoctoral fellowship from the National Institute of Health (ISCIII) of Spain.
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