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绿色荧光蛋白作为监测应激诱导型大鼠hsp70基因启动子活性的标记物。

Green fluorescent protein as a marker for monitoring activity of stress-inducible hsp70 rat gene promoter.

作者信息

Wysocka A, Krawczyk Z

机构信息

Department of Tumor Biology, Centre of Oncology-Maria Skłodowska-Curie Memorial Institute, Gliwice, Poland.

出版信息

Mol Cell Biochem. 2000 Dec;215(1-2):153-6. doi: 10.1023/a:1026523305294.

Abstract

Murine melanoma cells B16(F10) were stably transfected with a plasmid containing GFP gene linked to rat stress-inducible hsp70.1 gene promoter. Transfected cells show in vitro variable basal levels of fluorescence depending on stress response induced at physiological temperature by growth conditions. Lack of manipulations except medium change resulted in reduction of cellular fluorescence. GFP expression in experimental murine tumors dropped to levels undetectable at physiological temperature. Heat shock induced significant fluorescence of tumor cells both in vitro and in vivo. GFP protein could be a useful marker for studies of mammalian hsp70i gene promoters.

摘要

将含有与大鼠应激诱导型hsp70.1基因启动子相连的绿色荧光蛋白(GFP)基因的质粒稳定转染至小鼠黑色素瘤细胞B16(F10)。转染后的细胞在体外表现出不同的基础荧光水平,这取决于生理温度下生长条件所诱导的应激反应。除更换培养基外未进行其他操作会导致细胞荧光减弱。实验性小鼠肿瘤中的GFP表达在生理温度下降至无法检测的水平。热休克在体外和体内均能诱导肿瘤细胞产生显著荧光。GFP蛋白可能是研究哺乳动物hsp70i基因启动子的有用标志物。

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