Abstract
Porcine circovirus type 2 (PCV2) virus-like particles (VLPs) are an effective vaccine against post-weaning multisystemic wasting syndrome. Burst sequence (BS) and the chicken 5′-HS4 β-globin (HS4) insulator were used to improve the production of PCV2 VLPs in insect cells. BS was used to modify the polyhedron (polh) promoter, and the chicken HS4 insulator was inserted downstream of the target gene expression cassette. This enhanced effect was detected by EGFP, a generally used protein as reporter. The transcription-level analysis demonstrated that the double BS and the chicken HS4 insulator improved the transcription levels of the target genes. At the same time, the two regulatory elements also improved the PCV2 VLPs’ expression. PCV2 VLPs’ expression levels increased by 14 and 24 % at 72 h p.i. Further research observed an additive effect when the double BS and the chicken HS4 insulator simultaneously acted on the expression of PCV2 VLPs, with the PCV2 VLPs’ expression level increasing by 35 % at 72 h p.i. BS and HS4 insulator can be used to improve the production of PCV2 VLPs in insect cells.
Highlights
Porcine circovirus type 2 (PCV2) virus-like particles (VLPs) are an effective vaccine against post-weaning multisystemic wasting syndrome
Effects of the double Burst sequence (BS) and the chicken HS4 β-globin (HS4) insulator on enhanced green fluorescent protein (EGFP) protein expression Fluorescence can be observed by fluorescence microscope in all samples expect AcBlank-infected cells, indicating that AcBlank cannot express EGFP protein
The results showed that a 28 kDa band corresponding to the PCV2 Cap protein was present only in the AcCap-infected Spodopterafrugiperda cell line (Sf9) cells and not in the AcBlank-infected and non-infected Sf9 cells
Summary
Porcine circovirus type 2 (PCV2) virus-like particles (VLPs) are an effective vaccine against post-weaning multisystemic wasting syndrome. Burst sequence (BS) and the chicken 5′-HS4 β-globin (HS4) insulator were used to improve the production of PCV2 VLPs in insect cells. Porcine circovirus type 2, a member of the Circoviridae family, is a small, icosahedral, non-enveloped, singlestranded circovirus DNA virus (size ~17 nm) (Tischer et al 1982). The Cap protein is a major immunogen of PCV2. PCV2 is the primary causative agent of a group of diseases called porcine circovirus disease (PCVD) in swine. The most relevant diseases of PCVD are post-weaning multisystemic wasting syndrome (PMWS), porcine dermatitis and nephropathy syndrome (PDNS), and reproductive failure. PMWS has a serious economic impact on the world swine industry
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