Zhao Zhijing, Liu Xiumei
Institute of Nutrition and Food Hygiene, Chinese Academy of Preventive Medicine, Beijing 100050, China.
Wei Sheng Yan Jiu. 2002 Feb;31(1):49-52.
The green fluorescent protein (GFP) of jellyfish victoria is an unusual protein with strong visible fluorescence both in vivo and in vitro. In this study, about 750 bp fragment of gfp gene was amplified by PCR and was inserted into pRIT2T vector containing protein A gene. The recombinant plasmid was transferred into E. coli JM101. GFP and protein A were fused and expressed in E. coli Top10. The bright green fluorescence on the plates could be visible under UV at 365 nm. The bacteria cells were disrupted by sonic oscillation. The fused protein PA-GFP could be obtained from the supernatants.
维多利亚水母的绿色荧光蛋白(GFP)是一种不同寻常的蛋白质,在体内和体外均具有强烈的可见荧光。在本研究中,通过PCR扩增出约750 bp的gfp基因片段,并将其插入到含有蛋白A基因的pRIT2T载体中。重组质粒被转入大肠杆菌JM101。GFP与蛋白A融合并在大肠杆菌Top10中表达。平板上的亮绿色荧光在365 nm紫外线下可见。通过超声振荡破碎细菌细胞。可从上清液中获得融合蛋白PA-GFP。