Abstract:The characteristic properties of GFP make this protein a good candidate for use as a molecular reporter to monitor patterns of protein localization, gene expression, and intracellular protein trafficking in living cells. In this study, the dicistronic expression vector (pEGFP-C1) was used to be transfected into human lung cancer cell line(ASTC-a-1)and a positive clone which stably expressed GFP in high level was obtained. The results showed that the green fluorescent protein expressed in tumor cells was also photobleached under intense irradiation (-488 nm) and the degree of photobleaching varied with the intensity of the excitation. In order to analyze the effect of low temperature on the fluorescent sample, the photobleaching of tumor cell slice stored at -20 ℃ was observed additionally.
Key words:Green Fluorescent Protein (GFP);Human lung cancer cells;Photobleaching
金 鹰,邢 达*. 肿瘤细胞中表达的GFP蛋白的荧光漂白特性的研究[J]. 光谱学与光谱分析, 2004, 24(12): 1626-1629.
JIN Ying,XING Da*. Photobleaching Properties of GFP Expressed in Human Lung Cancer Cells . SPECTROSCOPY AND SPECTRAL ANALYSIS, 2004, 24(12): 1626-1629.
[1] Chalfie M, Tu Y, Euskirchen G et al. Science, 1994,263(5148): 802. [2] LI Bu-hong,XIE Shu-sen,LU Zu-kang(李步洪,谢树森,陆祖康). Chin. J. Laser Med. Surg.(中国激光医学杂志),2001,10(3): 151. [3] Wang S, Haxelrigg T. Nature, 1994,369: 400. [4] JIN Ying,XING Da(金 鹰,邢 达). Progress in Biochemistry and Biophysics(生物化学与生物物理进展),2002,29(6): 942. [5] Ogawa H, Inouye S et al. Prec. Natl. Acad. Sci. USA, 1995,92(25): 11899. [6] LI Nan,WANG Feng-xiang,ZHOU Chun-xi(李 南,王风翔,周春喜). Applied Technology of Fluorescent Probe(荧光探针应用技术). Beijing:Military and Medical Science Publishing Company(北京:军事医学科学出版社),1998. [7] Wang S, Hazelrigg T. Nature, 1994,369: 400. [8] Niswender K D, Blackman S M, Rohde L et al. J. Microbiol., 1995,180(2): 109. [9] YU Chang-qing, HUANG Nai-yan, ZHAO Hai-ying, DOU Xiao-ming, LI Jia-ze(于常青,黄乃艳,赵海鹰,窦晓鸣,李家泽). Spectroscopy and Spectral Analysis(光谱学与光谱分析),2004,24(2):138. [10] Lauf U, Giepmans B G. Prec. Natl. Acad. Sci. USA, 2002,99(16): 10446.