title: A green fluorescent protein-based reporter for protein nuclear import studies in Drosophila cells.
abstract: Green fluorescent protein-based reporters are commonly used to investigate protein nucleocytoplasmic transport. In this study we developed a novel reporter GFP2-GST which consists of two copies of GFP and one copy of GST, and tested it in two commonly used Drosophila cell lines. The size of the GFP2-GST reporter exceeds the passive diffusion limit across the nuclear pore complexes. It shows an exclusive cytoplasmic localization and displays a restrictive nuclear localization when a nuclear localization signal is appended. This reporter will largely facilitate the characterization and identification of NLS sequences in the fly proteome.
authors.affiliation: Laboratory of Drosophila Research, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China.
authors.forename: Wing Man
authors.initials: WM
authors.lastname: Chan
authors.forename: Pang Chui
authors.initials: PC
authors.lastname: Shaw
authors.forename: Ho Yin Edwin
authors.initials: HY
authors.lastname: Chan
mesh_terms: D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D002460:Cell Line; D004330:Drosophila / Q000235:genetics* / Q000378:metabolism*; D017930:Genes, Reporter; D005982:Glutathione Transferase / Q000737:chemistry / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics*; D019913:Nuclear Localization Signals; D022022:Nuclear Pore / Q000378:metabolism; D020543:Proteome; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics
chemical_list: D019913:Nuclear Localization Signals; D020543:Proteome; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D005982:Glutathione Transferase