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Title: Green fluorescent protein tagging: A novel tool in biomedical research
Authors: Akbar, Mohammed
Kim, Hee Yong
Keywords: plasma membrane;translocation;localization;PI3-kinase;Akt;PKC
Issue Date: Oct-2005
Publisher: CSIR
IPC Code: Int. Cl.7 G01N33/533; G03C1/73
Abstract: In recent years, the green fluorescent protein (GFP) isolated from the jellyfish, Aequorea victoria, has become a novel tool in biomedical research. The tagging of GFP to proteins of interest has widely been used in studies involving various biological events both in vitro and in vivo. In this report, the authors demonstrate the utility of GFP tagging for studying localization of protein kinase C (PKC ), and pleckstrin homology (PH) domains of phospholipase C (PLC1), and Akt in mouse neuroblastoma (Neuro 2A) cells. The authors also show the real time translocation of PKC and Akt-PH-EGFP to the membrane upon stimulation with either 12-phorbal myristate acetate (PMA) or insulin growth factor (IGF). Pretreatment of cells expressing PKC -EGFP or Akt-PH-EGFP with staurosporine (a protein kinase inhibitor) or wortmanin (a phosphatidylinositol 3-kinase inhibitor), prevented the translocation of these proteins to the membrane. These data demonstrate that, GFP tagging could be employed as a tool to study sub cellular localization and distribution of signaling proteins in response to external stimuli in real time.
Page(s): 466-470
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Appears in Collections:IJBT Vol.04(4) [October 2005]

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