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Coronaviruses pp 135-152 | Cite as

Engineering Infectious cDNAs of Coronavirus as Bacterial Artificial Chromosomes

  • Fernando Almazán
  • Silvia Márquez-Jurado
  • Aitor Nogales
  • Luis Enjuanes
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1282)

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.

Key words

Coronavirus MERS Reverse genetics Infectious clones Bacterial artificial chromosomes 

Notes

Acknowledgements

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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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Fernando Almazán
    • 1
  • Silvia Márquez-Jurado
    • 1
  • Aitor Nogales
    • 2
  • Luis Enjuanes
    • 1
  1. 1.Department of Molecular and Cell Biology, Centro Nacional de Biotecnología (CNB-CSIC)Campus Universidad Autónoma de MadridMadridSpain
  2. 2.Department of Microbiology and ImmunologyUniversity of RochesterRochesterUSA

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