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Real‐time bioluminescence imaging of a protein secretory pathway in living mammalian cells using Gaussia luciferase
Author(s) -
Suzuki Takahiro,
Usuda Shinsuke,
Ichinose Hiroshi,
Inouye Satoshi
Publication year - 2007
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2007.08.036
Subject(s) - luciferase , bioluminescence , chinese hamster ovary cell , luciferin , heterologous , bioluminescence imaging , microbiology and biotechnology , cell , chemistry , cell culture , biology , transfection , biochemistry , gene , genetics
Using photon counting and charge‐coupled device (CCD) cameras, we have applied the method of real‐time bioluminescence imaging to investigate protein trafficking in mammalian cells. In the living cells of Chinese hamster ovary and PC12D cells, exocytotic secretion of protein and protein targeting on the cell surface were visualized using the secreted Gaussia luciferase (GLase) as a reporter protein in a minute. After incubation of the cells with luciferin (coelenterazine) for 10 min, luciferin was imported into the cells and the vesicle transport network in the cells could be shown by luminescence images of GLase activity. Further, we demonstrate that GLase with a heterologous signal peptide sequence is targeted to the cell surface in neuronally differentiated PC12D cells and luminescence signals could be detected in a few seconds.