Viral 2A peptides allow expression of multiple proteins from a single ORF in transgenic zebrafish embryos

Authors

  • Elayne Provost,

    1. Department of Surgery, Johns Hopkins University School of Medicine, Baltimore, Maryland
    2. Department of Cell Biology, Johns Hopkins University School of Medicine, Baltimore, Maryland
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  • Jerry Rhee,

    1. Department of Surgery, Johns Hopkins University School of Medicine, Baltimore, Maryland
    2. Department of Cell Biology, Johns Hopkins University School of Medicine, Baltimore, Maryland
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  • Steven D. Leach

    Corresponding author
    1. Department of Surgery, Johns Hopkins University School of Medicine, Baltimore, Maryland
    2. Department of Cell Biology, Johns Hopkins University School of Medicine, Baltimore, Maryland
    • The Paul K. Neumann Professor in Pancreatic Cancer, Johns Hopkins University, Broadway Research Bldg., Rm 471, 733 N. Broadway, Baltimore, MD 21205
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Abstract

We have adapted a novel multicistronic gene expression system involving viral peptides to the zebrafish. The viral 2A peptide allows production of multiple protein products from a single transgene. Based on highly inefficient peptide bond formation between glycine and proline residues within the 2A peptide, placement of 2A peptide sequence as a linker region between tandem cDNA's allows the stoichiometric translation of multiple unfused protein products. To test this system in zebrafish, we generated two different tandem reporter constructs employing eGFP and mCherry, separated by 2A peptide sequence. Using this system, we produced transgenic zebrafish in which fluorophores were produced as independent proteins from a single transcript. The successful application of this technology in zebrafish will be valuable for visually marking transgenic embryos and transgene-expressing cells, or in any situation where reliable expression of multiple transgenes is desired. genesis 45:625–629, 2007. © 2007 Wiley-Liss, Inc.

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