High-efficiency expression of mammalian beta-adrenergic receptors in baculovirus-infected insect cells.
Infection of a clonal isolate of Spodoptera frugiperda cells (Sf9) with a baculovirus expression vector harboring the cDNA encoding the beta-adrenergic receptor resulted in a high efficiency expression. At 48 hr post-infection, the level of expression of beta-adrenergic receptors was approximately 12 million/cell. Specific activities of crude lysates of infected Sf9 cells were approximately 30 pmol/mg of protein, 5-fold greater than those of membranes of high-expressor Chinese hamster ovary cells stably transfected with an SV-40 expression vector. One liter of infected Sf9 cells expresses 20-40 nmol of receptor. Autoradiography of membranes incubated with the beta-adrenergic antagonist [125I]iodoazidobenzylpindolol, photolyzed, and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed 46,000- (presumably unglycosylated) and 48,000-Mr peptides for Sf9 cells as compared to approximately 65,000-Mr for Chinese hamster ovary cells. The baculovirus Sf9 system provides high-efficiency expression of receptor sufficient to permit physicochemical analyses.
George
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Virology Laboratory, Rockefeller University, New York, New York 10021.
Arbabian
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MA
Ruoho
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Kiely
J
J
Malbon
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CC
D000818:Animals; D002460:Cell Line; D002472:Cell Transformation, Viral; D006224:Cricetinae; D005796:Genes; D005822:Genetic Vectors; D007304:Insect Viruses / Q000235:genetics*; D007313:Insecta; D007700:Kinetics; D011943:Receptors, Adrenergic, beta / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
D011943:Receptors, Adrenergic, beta