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  Constitutive expression of recombinant human D2S-dopamine receptor in the unicellular yeast Saccharomyces cerevisiae

Sander, P., Grünewald, S., Maul, G., Reiländer, H., & Michel, H. (1994). Constitutive expression of recombinant human D2S-dopamine receptor in the unicellular yeast Saccharomyces cerevisiae. Biochimica et Biophysica Acta-Biomembranes, 1193(2), 255-262. doi:10.1016/0005-2736(94)90161-9.

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 Creators:
Sander, Peter1, Author           
Grünewald, Sylvia1, Author           
Maul, Gabi1, Author           
Reiländer, Helmut1, Author           
Michel, Hartmut1, Author                 
Affiliations:
1Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Max Planck Society, ou_2068290              

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Free keywords: Gene expression; Radioligand binding; D2S-dopamine receptor; Yeast
 Abstract: The cDNA for the human D2S-dopamine receptor has been functionally expressed in the unicellular yeast Saccharomyces cerevisiae. Two expression plasmids pRS421D2 (original D2S-gene coding region) and pRS421D2S (the first 24 aa of the yeast STE2-gene are fused to the N-terminus of the D2S-gene) were constructed and transformed into the protease deficient S. cerevisiae strain cI3-ABYS-86. Northern blot analysis of total RNA from transformed yeast clones revealed that for both constructs the D2S-gene was constitutively transcribed from the plasmids PMA1 promoter. Membranes prepared from recombinant S. cerevisiae exhibited saturable binding with the antagonist [3H]methylspiperone. Competition studies revealed pharmacological properties for these sites which were comparable to those reported for the D2-receptor heterologously expressed in mammalian cells. The expression of the receptor was monitored by Western blot analysis using an antiserum raised against a peptide from the third intracellular domain of the receptor protein and by ligand binding assay.

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Language(s): eng - English
 Dates: 1994-02-282002-12-191994-08-03
 Publication Status: Issued
 Pages: 8
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1016/0005-2736(94)90161-9
 Degree: -

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Title: Biochimica et Biophysica Acta-Biomembranes
Source Genre: Journal
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Publ. Info: Amsterdam : Elsevier
Pages: - Volume / Issue: 1193 (2) Sequence Number: - Start / End Page: 255 - 262 Identifier: ISSN: 0005-2736
CoNE: https://pure.mpg.de/cone/journals/resource/954926938702