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Coronavirus Replication and Reverse Genetics pp 229–252Cite as

Development of Mouse Hepatitis Virus and SARS-CoV Infectious cDNA Constructs

Part of the Current Topics in Microbiology and Immunology book series (CT MICROBIOLOGY,volume 287)

Abstract

The genomes of transmissible gastroenteritis virus (TGEV) and mouse hepatitis virus (MHV) have been generated with a novel construction strategy that allows for the assembly of very large RNA and DNA genomes from a panel of contiguous cDNA subclones. Recombinant viruses generated from these methods contained the appropriate marker mutations and replicated as efficiently as wild-type virus. The MHV cloning strategy can also be used to generate recombinant viruses that contain foreign genes or mutations at virtually any given nucleotide. MHV molecular viruses were engineered to express green fluorescent protein (GFP), demonstrating the feasibility of the systematic assembly approach to create recombinant viruses expressing foreign genes. The systematic assembly approach was used to develop an infectious clone of the newly identified human coronavirus, the serve acute respiratory syndrome virus (SARS-CoV). Our cloning and assembly strategy generated an infectious clone within 2 months of identification of the causative agent of SARS, providing a critical tool to study coronavirus pathogenesis and replication. The availability of coronavirus infectious cDNAs heralds a new era in coronavirus genetics and genomic applications, especially within the replicase proteins whose functions in replication and pathogenesis are virtually unknown.

Keywords

  • Recombinant Virus
  • Infectious Bronchitis Virus
  • Severe Acute Respiratory Syndrome
  • Infectious Clone
  • Mouse Hepatitis Virus

These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Authors and Affiliations

  1. Department of Epidemiology, University of North Carolina, Chapel Hill, NC, 27599-7400, USA

    R. S. Baric & A. C. Sims

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  1. R. S. Baric
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  2. A. C. Sims
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Editors and Affiliations

  1. Department of Molecular and Cell Biology, Centro Nacional de Biotecnología, Campus Universidad Autónoma, Cantoblanco, 38049, Madrid, Spain

    Professor Dr. Luis Enjuanes

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Baric, R.S., Sims, A.C. (2005). Development of Mouse Hepatitis Virus and SARS-CoV Infectious cDNA Constructs. In: Enjuanes, L. (eds) Coronavirus Replication and Reverse Genetics. Current Topics in Microbiology and Immunology, vol 287. Springer, Berlin, Heidelberg. https://doi.org/10.1007/3-540-26765-4_8

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