臨床検査51巻9号

2007年09月発行

今月の主題 フローサイトメトリー 技術・標識プローブ

GFP遺伝子の導入

著者： 長谷川洋¹ 小室一成¹

所属機関： ¹千葉大学大学院医学研究院循環病態医科学

ページ範囲：P.921 - P.926

　緑色蛍光蛋白質(GFP)は，発光クラゲから発見・分離された蛍光蛋白質であり，その安定性および簡便性から，レポーター分子として広く用いられており，遺伝子発現のマーカーや，様々な蛋白質の細胞内局在を調べるタグとしても用いられている．フローサイトメトリーを解析手段に用いることによって，その定量性は格段に向上し，再生医療や薬剤解析の網羅的解析等に威力を発揮している．

参考文献

１) Misteli T, Spector DL：Applications of the green fluorescent protein in cell biology and biotechnology. Nat Biotechnol　15：961-964, 1997

２) Shimomura O, Johnson FH, Saiga Y：Extraction, purification and properties of aequorin, a bioluminescent protein from the luminous hydromedusan, Aequorea. J Cell Comp Physiol　59：223-239, 1962

３) Shimomura O, Johnson FH：Chemical nature of bioluminescence systems in coelenterates. Proc Natl Acad Sci USA　72：1546-1549, 1975

４) Shimomura O：Isolation and properties of various molecular forms of aequorin. Biochem J　234：271-277, 1986

５) Prasher DC, Eckenrode VK, Ward WW, et al：Primary structure of the Aequorea victoria green-fluorescent protein. Gene　111：229-233, 1992

６) Arun KH, Kaul CL, Ramarao P：Green fluorescent proteins in receptor research：an emerging tool for drug discovery. J Pharmacol Toxicol Methods　51：1-23, 2005

７) Wang S, Hazelrigg T：Implications for bcd mRNA localization from spatial distribution of exu protein in Drosophila oogenesis. Nature　369：400-403, 1994

８) Hawley TS, Telford WG, Hawley RG：“Rainbow” reporters for multispectral marking and lineage analysis of hematopoietic stem cells. Stem Cells　19：118-124, 2001

９) Takada T, Iida K, Awaji T, et al：Selective production of transgenic mice using green fluorescent protein as a marker. Nat Biotechnol　15：458-461, 1997

10) Okabe M, Ikawa M, Kominami K, et al：'Green mice' as a source of ubiquitous green cells. FEBS Lett　407：313-319, 1997

11) Chishima T, Miyagi Y, Wang X, et al：Cancer invasion and micrometastasis visualized in live tissue by green fluorescent protein expression. Cancer Res　57：2042-2047, 1997

12) Iijima Y, Nagai T, Mizukami M, et al：Beating is necessary for transdifferentiation of skeletal muscle-derived cells into cardiomyocytes. FASEB J　17：1361-1363, 2003

13) Ohtsuka M, Takano H, Zou Y, et al：Cytokine therapy prevents left ventricular remodeling and dysfunction after myocardial infarction through neovascularization. FASEB J　18：851-853, 2004

14) Matsuura K, Wada H, Nagai T, et al：Cardiomyocytes fuse with surrounding noncardiomyocytes and reenter the cell cycle. J Cell Biol　167：351-363, 2004

15) Oyama T, Nagai T, Wada H, et al：Cardiac side population cells have a potential to migrate and differentiate into cardiomyocytes in vitro and in vivo. J Cell Biol　176：329-341, 2007

16) Niwa H, Miyazaki J, Smith AG：Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells. Nat Genet　24：372-376, 2000

17) Hidaka K, Lee JK, Kim HS, et al：Chamber-specific differentiation of Nkx2.5-positive cardiac precursor cells from murine embryonic stem cells. FASEB J　17：740-742, 2003

channel in mouse ES cell-derived cardiocytes carrying human chromosome 21. Biochem Biophys Res Commun　351：126-132, 2006

19) Terami H, Hidaka K, Shirai M, et al：Efficient capture of cardiogenesis-associated genes expressed in ES cells. Biochem Biophys Res Commun　355：47-53, 2007

20) Dell'Arciprete R, Stella M, Fornaro M, et al：High-efficiency expression gene cloning by flow cytometry. J Histochem Cytochem　44：629-640, 1996