Coronavirus Reverse Genetics by Targeted RNA Recombination
Targeted RNA recombination was the first reverse genetics system devised for coronaviruses at a time when it was not clear whether the construction of full-length infectious cDNA clones would become possible. In its current state targeted RNA recombination offers a versatile and powerful method for the site-directed mutagenesis of the downstream third of the coronavirus genome, which encodes all the viral structural proteins. The development of this system is described, with an emphasis on recent improvements, and multiple applications of this technique to the study of coronavirus molecular biology and pathogenesis are reviewed. Additionally, the relative strengths and limitations of targeted RNA recombination and infectious cDNA systems are contrasted.
KeywordsInfectious Bronchitis Virus Mouse Hepatitis Virus Reverse Genetic System Defective Interfere Murine Coronavirus
Unable to display preview. Download preview PDF.
- Casais R, Thiel V, Siddell SG, Cavanagh D, Britton P (2001) Reverse genetics system for the avian coronavirus infectious bronchitis virus. J Virol 75:12359–12369Google Scholar
- Nagy PD, Simon A (1997) New insights into the mechanisms of RNA recombination. Virology 235:1–9Google Scholar
- Rottier PJM (1995) The coronavirus membrane glycoprotein. In: Siddell SG (ed) The Coronaviridae. Plenum Press, New York, pp 115–139Google Scholar
- Vennema H, Poland A, Floyd-Hawkins K, Pedersen NC (1995) A comparison of the genomes of FECVs and FIPVs and what they tell us about the relationships between feline coronaviruses and their evolution. Feline Pract 23:40–44Google Scholar