High-level inducible expression of visual pigments in transfected cells.
Biotechniques
; 21(2): 304-11, 1996 Aug.
Article
em En
| MEDLINE
| ID: mdl-8862817
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 pMEP4 beta. The recombinant pMEP4 beta 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.
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Base de dados:
MEDLINE
Assunto principal:
Rodopsina
/
Proteínas Recombinantes de Fusão
/
Regulação da Expressão Gênica
/
Opsinas de Bastonetes
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Revista:
Biotechniques
Ano de publicação:
1996
Tipo de documento:
Article
País de afiliação:
Estados Unidos