<総説> 緑色蛍光蛋白質とフローサイトメトリーが拓くバイオテクノロジー研究の可能性

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In green fluorescent protein (GFP) is a protein extracted from the jellyfish, and produces the fluorescence similar to that of fluorescein isothiocyanate within living cells. Using flowcytometry, we can analyze and sort the cells expressing GFP fluorescence to accomplish the cellular and molecular biological experiments. We made the transgenic mice lines, and the knock-in mice, which had the GFP expression where endogenous GATA-2 gene was expressed. The GATA-2 is a transcription factor essential to the hematopoiesis, but its exact roles are not clarified yet. These mice produced the fluorescence in the hematopoietic and neural cells mimicking the transcriptional activity of the GATA-2 gene in individual cells. The fraction of the GFP expressing cells in the fetal liver, sorted by flowcytometry, had the high hematopoietic-colony-forming activities, indicating the GATA-2 expression in the hematopoietic progenitor cells. Further, we used the GFP to identify the cells infected by the retrovirus, and revealed the inhibitory role of GATA-2 to produce hematopoietic cells from their precursor mesenchymal cells. Thus, our experiments now heavily depend on the GFP, and the GFP techniques with flowcytometry would be the strong tool to reveal the physiological molecular mechanisms in the individual cells forming mammalian tissues.
筑波大学の論文