Publication

Reverse genetics of coronaviruses using vaccinia virus vectors

Book Chapter - Jan 1, 2005

Units
PubMed

Citation
Thiel V, Siddell S. Reverse genetics of coronaviruses using vaccinia virus vectors. In: Coronavirus Replication and Reverse Genetics. Heidelberg: Springer, 2005. ISBN 978-3-540-21494-6. pp. 199-227.
Type
Book Chapter (English)
Book Title
Coronavirus Replication and Reverse Genetics
Publication Date
Jan 1, 2005
Series Title
Current Topics in Microbiology and Immunology (287)
Isbn Number
978-3-540-21494-6
Pages
199-227
Publisher
Springer (Heidelberg)
Brief description/objective

In this article, we describe the reverse genetic system that is based on the use of vaccinia virus cloning vectors. This system represents a generic approach to coronavirus reverse genetics and was first described for the generation of recombinant human coronavirus 229E representing a group I coronavirus. Subsequently, the same approach has been used to generate recombinant avian infectious bronchitis coronavirus and, recently, recombinant mouse hepatitis virus, representing group III and group II coronaviruses, respectively. We describe how vaccinia virus-mediated homologous recombination can be used to introduce specific mutations into the coronavirus genomic cDNA during its propagation in vaccinia virus and how recombinant coronaviruses can be isolated. Finally, we describe how the coronavirus reverse genetic system has now been extended to the generation of coronavirus replicon RNAs.