Preparation of virus-like particles for porcine circovirus type 2 by YeastFab Assembly

Abstract

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.

Keywords: Capsid protein; Porcine circovirus type 2; Saccharomyces cerevisiae; Vaccine; Virus-like particle (VLP).