Journal
VIRUS GENES
Volume 53, Issue 4, Pages 661-666Publisher
SPRINGER
DOI: 10.1007/s11262-017-1457-8
Keywords
Influenza virus; Swine; 45 s rRNA promoter; Reverse genetics; Rescue
Categories
Funding
- National Natural Science Foundation of China [31572502]
- Agricultural Science and Technology Innovation Program (ASTIP)
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We generated an alternative reverse genetics (RG) system based on a synthesized swine 45S rRNA promoter to rescue the H3N2 subtype swine influenza virus. All eight flanking segment cassettes of A/swine/Henan/7/2010 (H3N2) were amplified with ambisense expression elements from RG plasmids. All segments were then recombined with the pHC2014 vector, which contained the synthesized swine 45S rRNA promoter (spol1) and its terminal sequence (t1) in a pcDNA3 backbone. As a result, we obtained a set of RG plasmids carrying the corresponding eight-segment cassettes. We efficiently generated the H3N2 virus after transfection into 293T/PK15, PK15, and 293T cells. The efficiency of spol1-driven influenza virus rescue in PK15 cells was similar to that in 293T cells by titration using the human pol1 RG system. Our approach suggests that an alternative spol1-based RG system can produce influenza viruses.
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