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 Scopus citations

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
DOIs
StatePublished - Aug 1996
Externally publishedYes

ASJC Scopus subject areas

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

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