Published in

Humana Press, Methods in Molecular Biology, p. 255-273, 2008

DOI: 10.1007/978-1-59745-181-9_19

Humana Press, Methods in Molecular Biology, p. 115-133, 2015

DOI: 10.1007/978-1-4939-2438-7_12

Humana Press, Methods in Molecular Biology, p. 147-165, 2020

DOI: 10.1007/978-1-0716-0900-2_12

Links

Tools

Export citation

Search in Google Scholar

Transient Dominant Selection for the Modification and Generation of Recombinant Infectious Bronchitis Coronaviruses

This paper is available in a repository.
This paper is available in a repository.

Full text: Download

Green circle
Preprint: archiving allowed
Green circle
Postprint: archiving allowed
Red circle
Published version: archiving forbidden
Data provided by SHERPA/RoMEO

Abstract

We have developed a reverse genetics system for the avian coronavirus infectious bronchitis virus (IBV) in which a full-length cDNA corresponding to the IBV genome is inserted into the vaccinia virus genome under the control of a T7 promoter sequence. Vaccinia virus as a vector for the full-length IBV cDNA has the advantage that modifications can be introduced into the IBV cDNA using homologous recombination, a method frequently used to insert and delete sequences from the vaccinia virus genome. Here, we describe the use of transient dominant selection as a method for introducing modifications into the IBV cDNA; this has been successfully used for the substitution of specific nucleotides, deletion of genomic regions, and the exchange of complete genes. Infectious recombinant IBVs are generated in situ following the transfection of vaccinia virus DNA, containing the modified IBV cDNA, into cells infected with a recombinant fowlpox virus expressing T7 DNA-dependent RNA polymerase.