Preparation of virus-like particles for porcine circovirus type 2 by YeastFab Assembly
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Porcine circovirus type 2 (PCV2) poses a genuine threat to pig industry. An effective vaccine production against the pandemic is desirable. The aim of this study was to construct recombination yeast strains with PCV2 Cap protein. We adopt to YeastFab Assembly method to synthesize transcriptional units in a single tube by piecing up promoter, open reading frame, and terminator in S. cerevisiae. Two yeast recombinants were successfully constructed using GPD and TEF2 promoters, respectively, to express PCV2 by secreting Cap protein in vitro. Electronic microscope observation demonstrated that the yeast-derived PCV2 Cap protein could self-assembles into 18-nm-diameter virus-like particles (VLPs). The yield of two different recombination yeasts containing GPD and TEF2 promoters were 12, 25 μg/ml, respectively. Our results showed that it is feasible to use S. cerevisiae as a safe and simple system to produce PCV2 virus-like particles. This indicated that there is possibility of obtaining PCV2 VLP vaccine by homologous recombination in yeast genome, and Cap protein was secreted into the cultural supernatant which can be used as a potential oral vaccine to protect pigs from PCV2-infection.
KeywordsSaccharomyces cerevisiae Porcine circovirus type 2 Capsid protein Virus-like particle (VLP) Vaccine
We thank prof. Junbiao Dai of Tsinghua University for providing us the YeastFab related materials and methods. This work was supported by the National Natural Science Foundation of Youth Science Foundation in China (31400064), and the Underprop Project of Tianjin Science and Technology Committee in China (16YFZCNC00640).
Conceived and designed the experiments: Jinhai Huang. Performed the experiments: Pei Chen, Tian Gao, Na Chang, Chengxue Zhao, Dong Lu, and Lilin Zhang. Analyzed the data: Lilin Zhang and Pei Chen. Contributed reagents/materials/analysis tools: Jinhai Huang. Wrote the paper: Pei Chen and Lilin Zhang.
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Conflict of interest
The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.
This article does not include any studies with human participants or animals performed by any of the authors.
Informed consent was obtained from all individual participants included in this article.