Impact of codon optimization and signal peptide on the expression of PCV2 ORF2 gene in Pichia pastoris
In order to efficiently express porcine circovirus 2(PCV2)ORF2 gene in Pichia pastoris,the codons of 187 amino acid of PCV2 ORF2 novel phytase gene was chemically synthesized by the PCR-based two-step DNA synthesis according to yeast codon preference.Unoptimized or optimized PCV2 ORF2 genes,with or without encoding signal peptide were inserted into pPICZα-C expression vector.The recombinant plasmids were linearized and transformed into Pichia pastoris GS115.Recombinant strains were preliminarily selected by Zeocin resistance.Only the PCV2 ORF2 gene,which was optimized gene and removed encoding signal peptide,was successfully expressed.The expression of recombinant protein was approved by SDS-PAGE and Western-blot analysis.The molecular weight of the target protein is about 30 ku.That the recombinant expressed protein could be recognized by anti-PCV2 serum which means the recombinant ORF2 protein had bioactivity.
Porcine circovirus 2(PCV2)ORF2 geneCodon optimizationRare codonNative signal peptidePichia pastoris
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