Construction of an infectious bronchitis virus vaccine strain carrying chimeric S1 gene of a virulent isolate and its pa
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APPLIED GENETICS AND MOLECULAR BIOTECHNOLOGY
Construction of an infectious bronchitis virus vaccine strain carrying chimeric S1 gene of a virulent isolate and its pathogenicity analysis Chenfei Lv 1 & Tingting Shi 1 & Pengpeng Zhu 1 & Xing Peng 1 & Shangshang Cao 1 & Yan Yan 1 & Nishant Kumar Ojha 1 & Min Liao 1 & Jiyong Zhou 1 Received: 28 March 2020 / Revised: 6 August 2020 / Accepted: 11 August 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Infectious bronchitis virus (IBV) is a member of genus gamma-coronavirus in the family Coronaviridae, causing serious economic losses to the poultry industry. Reverse genetics is a common technique to study the biological characteristics of viruses. So far, there is no BAC reverse genetic system available for rescue of IBV infectious clone. In the present study, a new strategy for the construction of IBV infectious cDNA clone was established. The full-length genomic cDNA of IBV vaccine strain H120 was constructed in pBAC vector from four IBV fragment subcloning vectors by homologous recombination, which contained the CMV promoter at the 5′ end and the hepatitis D virus ribozyme (HDVR) sequence and bovine growth hormone polyadenylation (BGH) sequence after the polyA tail at the 3′ end of the full-length cDNA. Subsequently, using the same technique, another plasmid pBAC-H120/SCS1 was also constructed, in which S1 gene from IBV H120 strain was replaced with that of a virulent SC021202 strain. Recombinant virus rH120 and rH120/SCS1 were rescued by transfecting the plasmids into BHK cells and passaged in embryonated chicken eggs. Finally, the pathogenicity of both the recombinant virus strains rH120 and rH120/SCS1 was evaluated in SPF chickens. The results showed that the chimeric rH120/SCS1 strain was not pathogenic compared with the wild-type IBV SC021202 strain and the chickens inoculated with rH120/SCS1 could resist challenge infection by IBV SC021202. Taken together, our results indicate that BAC reverse genetic system could be used to rescue IBV in vitro and IBV S1 protein alone might not be the key factor for IBV pathogenicity. Key points • BAC vector was used to construct IBV full-length cDNA by homologous recombination. • Based on four subcloning vectors, a recombinant chimeric IBV H120/SCS1 was constructed and rescued. • Pathogenicity of H120/SCS1 was similar to that of H120, but different to that of SC021202. Keywords Infectious bronchitis virus . Reverse genetics . Chimeric S1 gene . Pathogenicity
Introduction Infectious bronchitis virus (IBV) is a member of the Coronaviridae family in the order Nidovirales. It is a highly
Chenfei Lv and Tingting Shi contributed equally to this work. * Min Liao [email protected] 1
Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, People’s Republic of China
pathogenic virus that causes serious economic losses to the world poultry industry (Cavanagh 2005; Cook et al. 2012). IBV is an enveloped virus having a single-stranded, posi
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