Hyper-enhanced production of foreign recombinant protein by fusion with the partial polyhedrin of nucleopolyhedrovirus

To enhance the production efficiency of foreign protein in baculovirus expression systems, the effects of polyhedrin fragments were investigated by fusion expressing them with the enhanced green fluorescent protein (EGFP). Recombinant viruses were generated to express EGFP fused with polyhedrin frag...

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Published inPloS one Vol. 8; no. 4; p. e60835
Main Authors Bae, Sung Min, Kim, Hee Jung, Lee, Jun Beom, Choi, Jae Bang, Shin, Tae Young, Koo, Hyun Na, Choi, Jae Young, Lee, Kwang Sik, Je, Yeon Ho, Jin, Byung Rae, Yoo, Sung Sik, Woo, Soo Dong
Format Journal Article
LanguageEnglish
Published United States Public Library of Science 09.04.2013
Public Library of Science (PLoS)
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Summary:To enhance the production efficiency of foreign protein in baculovirus expression systems, the effects of polyhedrin fragments were investigated by fusion expressing them with the enhanced green fluorescent protein (EGFP). Recombinant viruses were generated to express EGFP fused with polyhedrin fragments based on the previously reported minimal region for self-assembly and the KRKK nuclear localization signal (NLS). Fusion expressions with polyhedrin amino acids 19 to 110 and 32 to 110 lead to localization of recombinant protein into the nucleus and mediate its assembly. The marked increase of EGFP by these fusion expressions was confirmed through protein and fluorescence intensity analyses. The importance of nuclear localization for enhanced production was shown by the mutation of the NLS within the fused polyhedrin fragment. In addition, when the polyhedrin fragment fused with EGFP was not localized in the nucleus, some fragments increased the production of protein. Among these fragments, some degradation of only the fused polyhedrin was observed in the fusion of amino acids 19 to 85 and 32 to 85. The fusion of amino acids 32 to 85 may be more useful for the enhanced and intact production of recombinant protein. The production of E2 protein, which is a major antigen of classical swine fever virus, was dramatically increased by fusion expression with polyhedrin amino acids 19 to 110, and its preliminary immunogenicity was verified using experimental guinea pigs. This study suggests a new option for higher expression of useful foreign recombinant protein by using the partial polyhedrin in baculovirus.
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Conceived and designed the experiments: SDW. Performed the experiments: SMB HJK JBL JBC TYS. Analyzed the data: HNK JYC KSL. Contributed reagents/materials/analysis tools: YHJ BRJ SSY. Wrote the paper: SMB SDW.
Competing Interests: Sung Sik Yoo is employed by Choong-Ang Vaccine Laboratory. There are no patents, products in development or marketed products to declare. This does not alter the authors’ adherence to all the PLOS ONE policies on sharing data and materials, as detailed online in the guide for authors.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0060835