+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

A reverse genetics system for avian coronavirus infectious bronchitis virus based on targeted RNA recombination



A reverse genetics system for avian coronavirus infectious bronchitis virus based on targeted RNA recombination



Virology Journal 14(1): 109



Avian coronavirus infectious bronchitis virus (IBV) is a respiratory pathogen of chickens that causes severe economic losses in the poultry industry worldwide. Major advances in the study of the molecular biology of IBV have resulted from the development of reverse genetics systems for the highly attenuated, cell culture-adapted, IBV strain Beaudette. However, most IBV strains, amongst them virulent field isolates, can only be propagated in embryonated chicken eggs, and not in continuous cell lines. We established a reverse genetics system for the IBV strain H52, based on targeted RNA recombination in a two-step process. First, a genomic and a chimeric synthetic, modified IBV RNA were co-transfected into non-susceptible cells to generate a recombinant chimeric murinized (m) IBV intermediate (mIBV). Herein, the genomic part coding for the spike glycoprotein ectodomain was replaced by that of the coronavirus mouse hepatitis virus (MHV), allowing for the selection and propagation of recombinant mIBV in murine cells. In the second step, mIBV was used as the recipient. To this end a recombination with synthetic RNA comprising the 3'-end of the IBV genome was performed by introducing the complete IBV spike gene, allowing for the rescue and selection of candidate recombinants in embryonated chicken eggs. Targeted RNA recombination allowed for the modification of the 3'-end of the IBV genome, encoding all structural and accessory genes. A wild-type recombinant IBV was constructed, containing several synonymous marker mutations. The in ovo growth kinetics and in vivo characteristics of the recombinant virus were similar to those of the parental IBV strain H52. Targeted RNA recombination allows for the generation of recombinant IBV strains that are not able to infect and propagate in continuous cell lines. The ability to introduce specific mutations holds promise for the development of rationally designed live-attenuated IBV vaccines and for studies into the biology of IBV in general.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 059398792

Download citation: RISBibTeXText

PMID: 28606144

DOI: 10.1186/s12985-017-0775-8


Related references

Reverse Genetics System for the Avian Coronavirus Infectious Bronchitis Virus. Methods in Molecular Biology 1602: 83-102, 2017

Reverse genetics system for the avian coronavirus infectious bronchitis virus. Journal of Virology 75(24): 12359-12369, 2001

Recombination in avian gamma-coronavirus infectious bronchitis virus. Viruses 3(9): 1777-1799, 2011

Sequence evidence for RNA recombination in field isolates of avian coronavirus infectious bronchitis virus. Vaccine 8(6): 605-608, 1990

Coronavirus proteins: Biogenesis of avian infectious bronchitis virus virion proteins. Structure and function of the oligosaccharides of the avian infectious bronchitis virus glycoproteins. Journal of Virology 44(3): 794-812, 1982

Use of defective RNAs containing reporter genes to investigate targeted recombination for avian infectious bronchitis virus. Advances in Experimental Medicine and Biology 494: 513-518, 2001

Reverse genetics of SARS-related coronavirus using vaccinia virus-based recombination. Plos one 7(3): E32857, 2012

Coronavirus reverse genetics by targeted RNA recombination. Current Topics in Microbiology and Immunology 287: 133-159, 2005

Hatchery Spray Cabinet Administration Does Not Damage Avian Coronavirus Infectious Bronchitis Virus Vaccine Based on Analysis by Electron Microscopy and Virus Titration. Avian Diseases 59(1): 149-152, 2015

Towards construction of viral vectors based on avian coronavirus infectious bronchitis virus for gene delivery and vaccine development. Journal of Virological Methods 160(1-2): 48-56, 2009

Coronavirus avian infectious bronchitis virus. Veterinary Research 38(2): 281-297, 2007

Experimental evidence of recombination in coronavirus infectious bronchitis virus. Virology 213(2): 569-580, 1995

Establishment of reverse genetics system for infectious bronchitis virus attenuated vaccine strain H120. Veterinary Microbiology 162(1): 53-61, 2013

Haemagglutination by avian infectious bronchitis virus-a coronavirus. Journal of General Virology 28(3): 381-390, 1975

Studies on the structure of a coronavirus-avian infectious bronchitis virus. Journal of General Virology 36(3): 495-502, 1977