Production of bovine rhodopsin by mammalian cell lines expressing cloned cDNA: Spectrophotometry and subcellular localization

Jeremy Nathans; Charles J. Weitz; Neeraj Agarwal; Izhak Nir; David S. Papermaster

doi:10.1016/0042-6989(89)90105-3

Production of bovine rhodopsin by mammalian cell lines expressing cloned cDNA: Spectrophotometry and subcellular localization

Jeremy Nathans, Charles J. Weitz, Neeraj Agarwal, Izhak Nir, David S. Papermaster

Howard Hughes Medical Institution

Research output: Contribution to journal › Article › peer-review

40 Scopus citations

Abstract

Cloned cDNA encoding bovine rhodopsin has been recombined into an expression vector and cotransfected with an antibiotic resistance plasmid into cultured human embryonic kidney cells. The resulting cell lines produce 100-200 μg of bovine opsin per liter of saturated tissue culture medium (10⁹ cells). Incubation in vitro with 11-cis retinal produces a photolabile pigment the absorbance spectrum of which is indistinguishable from that of bona fide bovine rhodopsin. Expressed rhodopsin accumulates in the plasma membrane as determined by immunoelectron microscopy.

Original language	English (US)
Pages (from-to)	907-914
Number of pages	8
Journal	Vision Research
Volume	29
Issue number	8
DOIs	https://doi.org/10.1016/0042-6989(89)90105-3
State	Published - 1989

Keywords

Immunoelectron microscopy
Transfection
Visual pigment

ASJC Scopus subject areas

Ophthalmology
Sensory Systems

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10.1016/0042-6989(89)90105-3

Cite this

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title = "Production of bovine rhodopsin by mammalian cell lines expressing cloned cDNA: Spectrophotometry and subcellular localization",

abstract = "Cloned cDNA encoding bovine rhodopsin has been recombined into an expression vector and cotransfected with an antibiotic resistance plasmid into cultured human embryonic kidney cells. The resulting cell lines produce 100-200 μg of bovine opsin per liter of saturated tissue culture medium (109 cells). Incubation in vitro with 11-cis retinal produces a photolabile pigment the absorbance spectrum of which is indistinguishable from that of bona fide bovine rhodopsin. Expressed rhodopsin accumulates in the plasma membrane as determined by immunoelectron microscopy.",

keywords = "Immunoelectron microscopy, Transfection, Visual pigment",

author = "Jeremy Nathans and Weitz, {Charles J.} and Neeraj Agarwal and Izhak Nir and Papermaster, {David S.}",

year = "1989",

doi = "10.1016/0042-6989(89)90105-3",

language = "English (US)",

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T1 - Production of bovine rhodopsin by mammalian cell lines expressing cloned cDNA

T2 - Spectrophotometry and subcellular localization

AU - Nathans, Jeremy

AU - Weitz, Charles J.

AU - Agarwal, Neeraj

AU - Nir, Izhak

AU - Papermaster, David S.

PY - 1989

Y1 - 1989

N2 - Cloned cDNA encoding bovine rhodopsin has been recombined into an expression vector and cotransfected with an antibiotic resistance plasmid into cultured human embryonic kidney cells. The resulting cell lines produce 100-200 μg of bovine opsin per liter of saturated tissue culture medium (109 cells). Incubation in vitro with 11-cis retinal produces a photolabile pigment the absorbance spectrum of which is indistinguishable from that of bona fide bovine rhodopsin. Expressed rhodopsin accumulates in the plasma membrane as determined by immunoelectron microscopy.

AB - Cloned cDNA encoding bovine rhodopsin has been recombined into an expression vector and cotransfected with an antibiotic resistance plasmid into cultured human embryonic kidney cells. The resulting cell lines produce 100-200 μg of bovine opsin per liter of saturated tissue culture medium (109 cells). Incubation in vitro with 11-cis retinal produces a photolabile pigment the absorbance spectrum of which is indistinguishable from that of bona fide bovine rhodopsin. Expressed rhodopsin accumulates in the plasma membrane as determined by immunoelectron microscopy.

KW - Immunoelectron microscopy

KW - Transfection

KW - Visual pigment

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JO - Vision Research

JF - Vision Research

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ER -

Production of bovine rhodopsin by mammalian cell lines expressing cloned cDNA: Spectrophotometry and subcellular localization

Abstract

Keywords

ASJC Scopus subject areas

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