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Expression of green fluorescent protein gene with baculovirus vector in insect cells
Authors:Hu Jianhong  Zhu Fanxiu  Qi Yipeng  Huang Yongxiu
Affiliation:(1) Institute of Virology, Wuhan University, 430072 Wuhan, China
Abstract:The green fluorescence of bioluminescent jellyfishAequorea victoria is due to the presence of the green fluorescent protein (GFP). To examine whether the GFP gene can be applied as a reporter gene in insect cells, a baculovirus transfer vector containing the neomycin resistance gene (neo) was established. The GFP gene was subcloned into the vector downstream of the polyhedrin gene (ocu) promoter. In the presence of G418, the recombinant virus can be purified. Expression of the GFP gene in the recombinant virus should give rise to synthesis of the GFP with a molecular weight of 30×103 dalton, and is observable by the strong green light irradiated by ultraviolet or blue light in viable intact insect cells. The GFP produced in insect cells has typical fluorescent spectra indistinguishable from those of the purified native GFP. The GFP gene as a good reporter gene can be applied to the baculovirus-insect cell expression system. Supported by the National Natural Science Foundation of China Hu Jianhong: born in July, 1972, Master graduate student
Keywords:green fluorescent protein (GFP)  baculovirus  transfer vector  insect cells  polyhedrin gene  neomycin resistance gene
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