[Expression of green fluorescent protein with baculovirus vector in insect cells].
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.
Zhu
F
F
Institute of Virology, Wuhan University.
Qi
Y
Y
Huang
Y
Y
Hu
J
J
D000818:Animals; D002478:Cells, Cultured; D004274:DNA, Recombinant / Q000096:biosynthesis; D015870:Gene Expression; D017930:Genes, Reporter; D005814:Genes, Viral; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics*; D009036:Moths / Q000166:cytology / Q000378:metabolism; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D014162:Transfection
D004274:DNA, Recombinant; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins