Fisheries and Aquatic Sciences

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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Functional Evaluation of the Rockbream (Oplegnathus fasciatus) Beta-actin Promoter as a Candidate Regulatory Element for DNA Vaccination

  • Kosuke, Zenke (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Lee, Sang-Yoon (Department of Aquaculture, Pukyong National University) ;
  • Kim, Ki-Hong (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Nam, Yoon-Kwon (Department of Aquaculture, Pukyong National University)
  • Published : 2009.06.30
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Abstract

The potential utility of the rockbream (Oplegnathus fasciatus) $\beta$-actin 5'-upstream sequence as a regulatory element for DNA vaccination was evaluated based on in vitro and in vivo heterologous expression assays. In the in vitro transfection experiment, the efficacy of the rockbream $\beta$-actin promoter to drive the expression of a downstream lacZ gene was significantly higher (more than fourfold) than that of the human cytomegalovirus (hCMV) promoter in two fish cell lines (grunt Haemulon plumierii fin and bluegill Lepomis macrochirus fry cell lines). In contrast, the functional activity of the rockbream $\beta$-actin promoter was hardly detectable in a mammalian mouse embryonic fibroblast cell line. Rockbream skeletal muscles injected in vivo with a GFP reporter construct driven by the $\beta$-actin promoter displayed the significantly higher expression of a GFP protein (more than threefold) than did those injected with hCMV promoter driven construct. Data from this study suggest that the homologous rockbream $\beta$-actin promoter could be used as a potential regulator for DNA vaccination in this species.

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References

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