Enhanced production of porcine circovirus type 2 (PCV2) virus-like particles in Sf9 cells by translational enhancers
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To investigate the effect of three translational enhancers for enhancing transgene expression in baculovirus expression vector system using GFP as a reporter gene and selected translational enhancers to increase porcine circovirus type 2 (PCV2) VLPs production.
P10UTR (the 3′-untranslated region from the baculovirus p10 gene), Syn21 (a synthetic AT-rich 21-bp sequence) and P10UTR/Syn21 increased the GFP yield by 1.4-, 4- and 4.8-fold, respectively. While IVS (intron from Drosophila myosin heavy chain gene) decreased the GFP yield by 65 %. Moreover, the synergy of P10UTR/Syn21 increased the yield of PCV2 VLPs by 4.1 fold (45 μg/106 cells) compared with standard baculovirus vector.
The synergy of P10UTR/Syn21 is a potential strategy to improve the recombinant vaccine production besides PCV2 VLPs in BEVS.
KeywordsBaculovirus Porcine circovirus type 2 Sf9 cell Translational enhancers Virus-like particles
This work was supported by National Natural Science Foundation of China (Grant No. 31101803/C1803) and Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (Grant No. SKLVBF201405).
Supplementary Table 1—Primers used in this study.
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