國家衛生研究院 NHRI:Item 3990099045/2754
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    題名: An ecdysone and tetracycline dual regulatory expression system for studies on Rac1 small GTPase-mediated signaling
    作者: Lai, JF;Juang, SH;Hung, YM;Cheng, HY;Cheng, TL;Mostov, KE;Jou, TS
    貢獻者: National Institute of Cancer Research
    摘要: Regulated expression systems are invaluable for studying gene function, offer advantages of dosage-dependent and temporally defined gene expression, and limit possible clonal variation when toxic or pleiotropic genes are overexpressed. Previously, establishment of inducible expression systems, such as tetracycline- and ecdysone-inducible systems, required assessment of the inducible characteristics of individual clones by tedious luciferase assays. Taking advantage of a green fluorescent protein (GFP) reporter controlled by tetracycline- or ecdysone-responsive element and fluorescence-activated cell sorting, we propose a simple and efficient strategy to select highly inducible cell lines according to their fluorescence profiles after transiently transfecting the candidate cell pools with a surrogate GFP reporter. We have demonstrated that tetracycline- and ecdysone-inducible systems could be set up in Madin-Darby canine kidney and HEK-293 cells by employing this selection scheme. Importantly, this dual regulatory expression system is applied in studying the complex interplay between two Ras-related small GTPases, Cdc42 and Rac1, on detachment-induced apoptosis. Furthermore, establishment of two tightly regulated expression systems in one target cell line could be of great advantage for dissecting small GTPase Rac1-transduced signaling pathways by using global gene expression approaches such as proteomic assays.
    關鍵詞: Cell Biology;Physiology
    日期: 2003-09
    關聯: American Journal of Physiology-Cell Physiology. 2003 Sep;285(3):C711-C719.
    Link to: http://ajpcell.physiology.org/cgi/content/full/285/3/C711
    JIF/Ranking 2023: http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=NHRI&SrcApp=NHRI_IR&KeyISSN=0363-6143&DestApp=IC2JCR
    Cited Times(WOS): https://www.webofscience.com/wos/woscc/full-record/WOS:000184549600024
    Cited Times(Scopus): http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=0042388528
    顯示於類別:[莊聲宏(1997-2004)] 期刊論文

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