Open AccessExpression of a bovine rhodopsin gene in Xenopus oocytes: demonstration of light-dependent ionic currents.
Author(s) -
H. G. Khorana,
Barry E. Knox,
Enrico Nasi,
Richard Swanson,
Debra A. Thompson
Publication year - 1988
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.85.21.7917
Subject(s) - rhodopsin , xenopus , opsin , biology , microbiology and biotechnology , gtp' , voltage clamp , messenger rna , gene expression , visual phototransduction , retinal , gene , biochemistry , biophysics , membrane potential , enzyme
Xenopus oocytes express a gene encoding bovine rhodopsin as well as its SP6 RNA polymerase-derived transcripts and total retinal mRNA. The opsin produced is in unglycosylated (30 kDa) and two glycosylated (35 kDa and 41 kDa) forms. Incubation of the cells expressing the above proteins with 11-cis-retinal generates rhodopsin, which was purified by immunoaffinity chromatography. The purified protein shows the expected UV/visible absorption spectrum and characteristic light-dependent activation of the rod outer segment GTP binding protein. Oocytes expressing rhodopsin exhibit light-dependent ionic currents that are detected by voltage-clamp techniques.