TY - JOUR T1 - Construction of a Chimeric Virus Expressing Mutation Sequences of Classical Swine Fever Virus Yunnan Strain AU - Yang, Yu-Ai AU - Sun, Yong-Ke AU - Zhang, Xiao-Min AU - Du, Min AU - Li, Yu-Xin AU - Kong, Ling-Fu AU - Pan, Hong-Bin AU - Yan, Yu-Lin JO - Journal of Animal and Veterinary Advances VL - 13 IS - 7 SP - 470 EP - 476 PY - 2014 DA - 2001/08/19 SN - 1680-5593 DO - javaa.2014.470.476 UR - https://makhillpublications.co/view-article.php?doi=javaa.2014.470.476 KW - RNA KW -CSFV KW -chimeric virus KW -construction KW -genetic stability KW -infectious molecular clone AB - 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. ER -