Journal of Animal and Veterinary Advances
ISSN: Online 1993-601XISSN: Print 1680-5593
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Construction of a Chimeric Virus Expressing Mutation Sequences of Classical Swine Fever Virus Yunnan Strain
Yong-Ke Sun, Xiao-Min Zhang, Min Du, Yu-Xin Li, Ling-Fu Kong, Hong-Bin Pan, Yu-Lin Yan and Yu-Ai Yang
Page: 470-476 | Received 21 Sep 2022, Published online: 21 Sep 2022
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Abstract
Infectious cDNA clones are a prerequisite for directed genetic manipulation of RNA viruses. To explore the role of mutations in Classical Swine Fever Virus (CSFV) Yunnan strain which caused the atypical clinical signs in pigs, a new pSM derived from CSFV Shiman strain has been constructed and subsequently replaced by the mutation sequences of CSFV YN strain isolated by the laboratory at the positions 1510-1532, 2471-2658, 3152-3176 and 11785-11816 using the targeted recombination strategy to enable rescue of chimeric CSFV. The results showed that chimeric CSFV (vSM-YN) was successfully rescued from PK-15 cells by transfection of the chimeric CSFV RNA transcripts and identified by whole genome sequence analysis, immunofluorescence antibody assay and ELISA detection. Sequencing of the pAC-SM-YN revealed a high genetic stability and the complete genome sequences of rescued viruses vSM-YN after extensive passages in PK-15 cells showed that modifications in pSM were stably maintained. The results indicate that targeted recombination-mediated mutagenesis provides a powerful tool for expediting the construction of novel RNA genomes and should facilitate further study of the pathogenic mechanism of CSFV leading to atypical CSF.
How to cite this article:
Yong-Ke Sun, Xiao-Min Zhang, Min Du, Yu-Xin Li, Ling-Fu Kong, Hong-Bin Pan, Yu-Lin Yan and Yu-Ai Yang. Construction of a Chimeric Virus Expressing Mutation Sequences of Classical Swine Fever Virus Yunnan Strain.
DOI: https://doi.org/10.36478/javaa.2014.470.476
URL: https://www.makhillpublications.co/view-article/1680-5593/javaa.2014.470.476