Zhu F, Qi Y, Huang Y, Hu J
Institute of Virology, Wuhan University.
Wei Sheng Wu Xue Bao. 1997 Feb;37(1):15-20.
The green fluorescent protein (GFP) gene was subcloned into the transfer vector pVLneo downstream of the polyhedrin gene (ocu) promoter. Insect cells were cotransfected with recombinant plasmid and Autographa californica Nuclear Polyhedrosis Virus (AcNPV) DNA. In the presence of G418, the recombinant virus containing GFP gene was purified. The GFP expressed in insect cells with a Mw of 30 kDa is observable by strong green light under a fluorescent microscope. Excitation and emission spectra of the GFP were 395 nm and 509 nm respectively. Integration of GFP gene on AcNPV genome was identified directly by Southern blot which gave strong hybridization signal between GFP cDNA probe and 1 kb EcoRI fragment of recombinant virus.
绿色荧光蛋白（GFP）基因被亚克隆到多角体蛋白基因（ocu）启动子下游的转移载体pVLneo中。昆虫细胞用重组质粒和苜蓿银纹夜蛾核型多角体病毒（AcNPV）DNA共转染。在G418存在的情况下，纯化含有GFP基因的重组病毒。在荧光显微镜下，通过强绿光可观察到在昆虫细胞中表达的分子量为30 kDa的GFP。GFP的激发光谱和发射光谱分别为395 nm和509 nm。通过Southern印迹直接鉴定GFP基因在AcNPV基因组上的整合，该印迹在GFP cDNA探针与重组病毒的1 kb EcoRI片段之间产生强杂交信号。
