A green fluorescent protein-based reporter for protein nuclear import studies in Drosophila cells.
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.
Chan
Wing Man
WM
Laboratory of Drosophila Research, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong, China.
Shaw
Pang Chui
PC
Chan
Ho Yin Edwin
HY
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
D019913:Nuclear Localization Signals; D020543:Proteome; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D005982:Glutathione Transferase