Efficient Production of Porcine Circovirus Type 2 Capsid Protein using Baculovirus

Abstract

Porcine circovirus type 2 (PCV2) is a single-stranded circular DNA virus associated with Postweaning multisystemic wasting syndrome (PMWS), which is considered to be an important infectious swine viral disease. PCV2 capsid protein encoded by ORF2 is a structural protein and expected as the high immunogenicity protein. In this study, we generated recombinant baculovirus containing ORF2 of PCV2 and analyzed the optimal conditions for the production of capsid protein in insect cell. Production and status of recombinant capsid protein in insect cell were confirmed by SDS-PAGE and Western blot analysis using His tag antibody and anti-PCV2 serum. The yield of recombinant capsid protein was high like as shown visible on SDS-PAGE. Optimal multiplicity of infection (MOI) and infection time of recombinant virus were determined as 5 MOI and 4 days, respectively. ORF2 is known to have Nlinked glycosylation site, but we couldn't detect the glycosylation of recombinant protein in insect cells.

Expression of porcine circovirus type 2 capsid protein fused with partial polyhedrin using baculovirus

“…In our previous report, we did not observe the glycosylation of recombinant PCV2 capsid proteins in insect cells (Lee et al, 2012). The glycosylation process is important in the posttranslational modification of proteins because it may confer activity to the proteins.…”

“…The previously constructed pB9-PCV2ORF2 (Lee et al, 2012) containing ORF2 gene was used in this study. The nuhost .…”

“…Previously, we reported that the BEVS is sufficient to production of PCV2 capsid protein (Lee et al, 2012). In this study, we generated recombinant baculoviruses to express …”