Abstract：In order to produce HPV16 L1 protein and self-assemble virus-like particles (VLPs), optimized HPV16 L1 gene according to the yeast cell codon preference was synthesized and cloned into pPIC3.5K yeast expression vector. After linearized with Bgl II restriction enzyme, pPIC3.5K/HPV16 L1 recombinant plasmid was electricity transformed into GS115 strain to construct HPV16 L1-recombinant Pichia pastoris. Identified recombinant strains were then induced by methanol. The expression of HPV16 L1 was identified by monoclonal antibodies, and self-assemble VLPs was purified by heparin and observed under transmission electron microscope. Data of PCR detection, restriction enzyme digestion and sequencing analysis showed that pPIC3.5 K/HPV16 L1 recombinant plasmid was successfully constructed. Western blot verify the expression of HPV16 L1 in the lysate of HPV16 L1 recombinant Pichia pastoris. After purified by Heparin, VLPs with diameter of about 55 nm were observed in the transmission electron microscopy. Their shapes were similar to natural HPV16 virus particles. In conclusion, we successfully expressed HPV16 L1 protein in integrated recombinant Pichia pastoris system and quickly obtained intact conformation of VLPs by using heparin affinity chromatography. This study will facilitate the development of HPV16 prophylactic vaccine.
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2014, Vol. 30 
