High-level inducible expression of visual pigments in transfected cells

Manija A. Kazmi, Robert A. Dubin, Carole Oddoux, Harry Ostrer

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

A method for high-level expression of a functionally active, recombinant human red cone opsin was developed by adding the coding sequence for the C-terminal epitope of bovine rhodopsin onto the C terminus of the cone opsin and cloning the resulting construct into the vector pMEP4β. The recombinant pMEP4β vector was transfected stably into 293-EBNA cells, and expression of the cone opsin was induced by the addition of CdCl2 into the medium. The recombinant cone opsin was reconstituted with 11-cis retinal and purified by immunoaffinity chromatography. Spectral analysis prior to and following photobleaching confirmed its identity as a red cone opsin. The protein was targeted to the cell membrane and activated bovine transducin.

Original languageEnglish (US)
Pages (from-to)304-311
Number of pages8
JournalBioTechniques
Volume21
Issue number2
StatePublished - Aug 1996
Externally publishedYes

Fingerprint

Cone Opsins
Retinal Pigments
Transducin
Retinaldehyde
Photobleaching
Cadmium Chloride
Rhodopsin
Cloning
Cell membranes
Chromatography
Spectrum analysis
Organism Cloning
Epitopes
Cell Membrane
Proteins

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Kazmi, M. A., Dubin, R. A., Oddoux, C., & Ostrer, H. (1996). High-level inducible expression of visual pigments in transfected cells. BioTechniques, 21(2), 304-311.

High-level inducible expression of visual pigments in transfected cells. / Kazmi, Manija A.; Dubin, Robert A.; Oddoux, Carole; Ostrer, Harry.

In: BioTechniques, Vol. 21, No. 2, 08.1996, p. 304-311.

Research output: Contribution to journalArticle

Kazmi, MA, Dubin, RA, Oddoux, C & Ostrer, H 1996, 'High-level inducible expression of visual pigments in transfected cells', BioTechniques, vol. 21, no. 2, pp. 304-311.
Kazmi, Manija A. ; Dubin, Robert A. ; Oddoux, Carole ; Ostrer, Harry. / High-level inducible expression of visual pigments in transfected cells. In: BioTechniques. 1996 ; Vol. 21, No. 2. pp. 304-311.
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