Codon optimization for expression of porcine circovirus ORF2 protein in Escherichia coli

Abstract

In this paper, some results of codon optimization of porcine circovirus ORF2 gene and expression of ORF2 protein in Escherichia coli were presented. Optimal ORF2 gene was successfully cloned in expression vector pET22b (+) and transformed into E. coli BL21(DE3) host. Expression of ORF2 protein in E. coli BL21(DE3) was checked by polyacrylamide gel electrophoresis and Western blot method. Optimization studies of expression conditions for recombinant strain indicated that the highest level of recombinant ORF2 was achieved in conditions of 0.05% isopropyl β-D-1-thiogalactopyranoside (IPTG) inducer, 37oC temperature after 4 hours induction. Recombinant ORF2 protein was purified using His-tag specific affinity chromatography and could be used as antigen for animal immunization to generate specific antibodies. Successful expression of porcine circovirus ORF2 protein in E. coli BL21(DE3) could provide antigen source for applied studies in diagnosis and development of novel vaccines.