Enhanced production of porcine circovirus type 2 (PCV2) virus-like particles in Sf9 cells by translational enhancers
- 653 Downloads
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.
- Royall E, Woolaway KE, Schacherl J, Kubick S, Belsham GJ, Roberts LO (2004) The Rhopalosiphum padi virus 5′ internal ribosome entry site is functional in Spodoptera frugiperda 21 cells and in their cell-free lysates: implications for the baculovirus expression system. J Gen Virol 85:1565–1569CrossRefPubMedGoogle Scholar