Zhao, Ye; Cheng, Jinlong; Xu, Gang; Thiel, Volker; Zhang, Guozhong (2019). Successful establishment of a reverse genetic system for QX-type infectious bronchitis virus and technical improvement of the rescue procedure. Virus research, 272, p. 197726. Elsevier 10.1016/j.virusres.2019.197726
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In this study, a pathogenic avian infectious bronchitis virus (IBV) QX-type strain YN was successfully rescued by vaccinia virus based reverse genetic technology. Ten fragments contiguously spanning the complete IBV genome were amplified and cloned into the vaccinia virus genome by homologous recombination. The full-length genomic cDNA was transcribed in vitro, and its transcript was transfected into BHK-21/N cells that could stably express IBV N protein. At 48 h post transfection, the culture medium was harvested and inoculated into 10-day-old specific-pathogen-free embryonated chicken eggs to replicate the rescued virus. This strategy was chosen to facilitate the rescue procedure and to ensure that the recombinant rYN virus will not require any cell culture adaptations. After only one in ovo passage, the recombinant YN virus (rYN) was successfully recovered and confirmed to possess the introduced silent marker mutation in its genome. Biological characteristics of rYN such as the EID50, TCID50, replication in ovo, and replication kinetcs in vitro were tested and all were similar to its parental strain YN. Our findings demonstrate the successful construction of highly-pathogenic QX-type IBV using a modified rescue procedure, allowing for future studies of the molecular biology and pathogenicity of IBV field strains.
Item Type: |
Journal Article (Original Article) |
|---|---|
Division/Institute: |
05 Veterinary Medicine > Research Foci > Host-Pathogen Interaction 05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Virology and Immunology 05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) |
UniBE Contributor: |
Thiel, Volker Earl |
Subjects: |
500 Science > 570 Life sciences; biology 600 Technology > 610 Medicine & health 600 Technology > 630 Agriculture |
ISSN: |
0168-1702 |
Publisher: |
Elsevier |
Language: |
English |
Submitter: |
Pamela Schumacher |
Date Deposited: |
09 Oct 2019 12:25 |
Last Modified: |
05 Dec 2022 15:31 |
Publisher DOI: |
10.1016/j.virusres.2019.197726 |
PubMed ID: |
31430502 |
Uncontrolled Keywords: |
Coronavirus IBV QX-type Reverse genetic system Vaccinia virus |
BORIS DOI: |
10.7892/boris.133782 |
URI: |
https://boris.unibe.ch/id/eprint/133782 |
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