Development of an improved dual-promoter-based reverse genetics system for emerging Senecavirus A: Running title: A reverse genetics system for Senecavirus A.

Development of an improved dual-promoter-based reverse genetics system for emerging Senecavirus A: Running title: A reverse genetics system for Senecavirus A. J Virol Methods. 2020 Sep 14;:113973 Authors: Zhang X, Lu J, Deng T, Zhao P, Peng Z, Chen L, Qian M, Guo Y, Qiao H, Wang Y, Xia Y, Bian C, Wang Z Abstract Senecavirus A (SVA), a recently emerging picornavirus, poses a great threat to the swine industry because it causes swine idiopathic vesicular disease and epidemic transient neonatal losses. Thus far, the progress in SVA viral pathogenesis studies and vaccine development remains sluggish, and an available and convenient reverse genetics system would undoubtedly promote relevant research. Herein, we established an improved universal dual-promoter reverse genetics system with an SVA-specific hammerhead ribozyme and hepatitis delta virus ribozyme at both terminals of the viral genome; this system could be applied to rescue all SVA strains by both eukaryotic and prokaryotic RNA polymerase systems. The genome of the clone-derived Chinese field strain CH/HeN-2018 was assembled into the universal vector pcDNA-rSVAuni through the Gibson assembly technique. Moreover, two silent mutations, G6848C and C7163 G, were separately engineered into the full-length cDNA clone with one step site-directed mutagenesis to create a KpnI restriction enzyme site, which served as a unique genetic marker. The viruses, designated rCH/HeN-2018-T7, rCH/H...
Source: Journal of Virological Methods - Category: Virology Authors: Tags: J Virol Methods Source Type: research