High-efficiency expression of mammalian beta-adrenergic receptors in baculovirus-infected insect cells. 1989

S T George, and M A Arbabian, and A E Ruoho, and J Kiely, and C C Malbon
Virology Laboratory, Rockefeller University, New York, New York 10021.

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.

UI MeSH Term Description Entries
D007304 Insect Viruses Viruses infecting insects, the largest family being BACULOVIRIDAE. Insect Virus,Virus, Insect,Viruses, Insect
D007313 Insecta Members of the phylum ARTHROPODA composed or organisms characterized by division into three parts: head, thorax, and abdomen. They are the dominant group of animals on earth with several hundred thousand different kinds. Three orders, HEMIPTERA; DIPTERA; and SIPHONAPTERA; are of medical interest in that they cause disease in humans and animals. (From Borror et al., An Introduction to the Study of Insects, 4th ed, p1). Insects,Insect
D007700 Kinetics The rate dynamics in chemical or physical systems.
D011943 Receptors, Adrenergic, beta One of two major pharmacologically defined classes of adrenergic receptors. The beta adrenergic receptors play an important role in regulating CARDIAC MUSCLE contraction, SMOOTH MUSCLE relaxation, and GLYCOGENOLYSIS. Adrenergic beta-Receptor,Adrenergic beta-Receptors,Receptors, beta-Adrenergic,beta Adrenergic Receptor,beta-Adrenergic Receptor,beta-Adrenergic Receptors,Receptor, Adrenergic, beta,Adrenergic Receptor, beta,Adrenergic beta Receptor,Adrenergic beta Receptors,Receptor, beta Adrenergic,Receptor, beta-Adrenergic,Receptors, beta Adrenergic,beta Adrenergic Receptors,beta-Receptor, Adrenergic,beta-Receptors, Adrenergic
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D002472 Cell Transformation, Viral An inheritable change in cells manifested by changes in cell division and growth and alterations in cell surface properties. It is induced by infection with a transforming virus. Transformation, Viral Cell,Viral Cell Transformation,Cell Transformations, Viral,Transformations, Viral Cell,Viral Cell Transformations
D005796 Genes A category of nucleic acid sequences that function as units of heredity and which code for the basic instructions for the development, reproduction, and maintenance of organisms. Cistron,Gene,Genetic Materials,Cistrons,Genetic Material,Material, Genetic,Materials, Genetic
D005822 Genetic Vectors DNA molecules capable of autonomous replication within a host cell and into which other DNA sequences can be inserted and thus amplified. Many are derived from PLASMIDS; BACTERIOPHAGES; or VIRUSES. They are used for transporting foreign genes into recipient cells. Genetic vectors possess a functional replicator site and contain GENETIC MARKERS to facilitate their selective recognition. Cloning Vectors,Shuttle Vectors,Vectors, Genetic,Cloning Vector,Genetic Vector,Shuttle Vector,Vector, Cloning,Vector, Genetic,Vector, Shuttle,Vectors, Cloning,Vectors, Shuttle
D006224 Cricetinae A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS. Cricetus,Hamsters,Hamster
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia

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