Expression of the gene encoding firefly luciferase in insect cells using a baculovirus vector.
A cDNA encoding the firefly luciferase [Photinus luciferin: oxygen 4-oxidoreductase (decarboxylating, ATP-hydrolyzing), EC 1.13.12.7] was cloned downstream from the polyhedrin gene promoter of Autographa californica nuclear polyhedrosis virus and expressed in Spodoptera frugiperda clone-9 cells. Synthesis of luciferase (Luc) was accurately measured in insect cells growing in a 96-well plate, by a simple, rapid, nonradioactive, inexpensive and sensitive method based on fogging of x-ray film. Luc was produced in a coordinate fashion during virus infection. The Luc synthesized in insect cells was not secreted into the medium but was contained within the cell. Our findings suggest that Luc can be used as a superior reporter enzyme for molecular genetic analyses of baculovirus regulatory signals involved in high level expression of foreign genes, protein processing, targeting and stability in insect cells.
National Institute of Immunology, New Delhi, India.
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Hasnain
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Nakhai
D000818:Animals; D002460:Cell Line; D001517:Coleoptera / Q000201:enzymology / Q000235:genetics*; D015870:Gene Expression; D005796:Genes; D007304:Insect Viruses / Q000235:genetics*; D007313:Insecta; D007700:Kinetics; D008156:Luciferases / Q000235:genetics* / Q000378:metabolism; D015183:Restriction Mapping; D014162:Transfection
D008156:Luciferases