Enhanced production of porcine circovirus type 2 (PCV2) virus-like particles in Sf9 cells by translational enhancers

OBJECTIVE: 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. RESULTS: P10UTR (the 3′-unt...

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Bibliographic Details
Published in:Biotechnology letters 2015-09, Vol.37 (9), p.1765-1771
Main Authors: Liu, Yangkun, Zhang, Yuanyuan, Yao, Lunguang, Hao, Huafang, Fu, Xiangjing, Yang, Zengqi, Du, Enqi
Format: Article
Language:English
Subjects:
Animal diseases
Animals
Applied Microbiology
Baculoviridae - genetics
Baculoviridae - metabolism
Baculovirus
Biochemistry
Biomedical and Life Sciences
Biotechnology
Capsid Proteins - genetics
Capsid Proteins - metabolism
Circovirus - genetics
Circovirus - metabolism
Drosophila
Enhancer Elements, Genetic
Gene expression
Genes
Green Fluorescent Proteins - genetics
Green Fluorescent Proteins - metabolism
Hogs
introns
Life Sciences
Mathematical analysis
Microbiology
myosin heavy chains
Original Research Paper
Porcine circovirus
Porcine circovirus-2
Recombinant
recombinant vaccines
reporter genes
Sf9 Cells
Strategy
Swine
translation (genetics)
Vaccines
Vaccines, Virus-Like Particle - genetics
Vaccines, Virus-Like Particle - metabolism
Vectors (mathematics)
Veterinary medicine
Viral Proteins - genetics
virus-like particles
Viruses
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Summary:OBJECTIVE: 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. RESULTS: 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. CONCLUSION: The synergy of P10UTR/Syn21 is a potential strategy to improve the recombinant vaccine production besides PCV2 VLPs in BEVS.
ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-015-1856-7