title: Viral 2A peptides allow expression of multiple proteins from a single ORF in transgenic zebrafish embryos.
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.
authors.affiliation: Department of Surgery, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
authors.forename: Elayne
authors.initials: E
authors.lastname: Provost
authors.forename: Jerry
authors.initials: J
authors.lastname: Rhee
authors.forename: Steven D
authors.initials: SD
authors.lastname: Leach
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D018076:DNA, Complementary; D004625:Embryo, Nonmammalian; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008164:Luminescent Proteins / Q000378:metabolism; D016366:Open Reading Frames; D010455:Peptides / Q000235:genetics*; D019076:Transgenes; D015027:Zebrafish / Q000196:embryology / Q000235:genetics*; D029961:Zebrafish Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D008164:Luminescent Proteins; D010455:Peptides; D029961:Zebrafish Proteins; D049452:Green Fluorescent Proteins