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Titolo:
Evaluation of the tetracycline- and ecdysone-inducible systems for expression of neurotransmitter receptors in mammalian cells
Autore:
Van Craenenbroeck, K; Vanhoenacker, P; Leysen, JE; Haegeman, G;
Indirizzi:
State Univ Ghent, Dept Mol Biol, B-9000 Ghent, Belgium State Univ Ghent Ghent Belgium B-9000 pt Mol Biol, B-9000 Ghent, Belgium Janssen Res Fdn, B-2340 Beerse, Belgium Janssen Res Fdn Beerse Belgium B-2340 en Res Fdn, B-2340 Beerse, Belgium
Titolo Testata:
EUROPEAN JOURNAL OF NEUROSCIENCE
fascicolo: 6, volume: 14, anno: 2001,
pagine: 968 - 976
SICI:
0953-816X(200109)14:6<968:EOTTAE>2.0.ZU;2-L
Fonte:
ISI
Lingua:
ENG
Soggetto:
CONTROLLED GENE-EXPRESSION; TUMOR-NECROSIS-FACTOR; RETINOID-X-RECEPTOR; EPSTEIN-BARR-VIRUS; TRANSGENIC MICE; RESPONSIVE PROMOTER; TEMPORAL CONTROL; NITRIC-OXIDE; IN-VIVO; DROSOPHILA;
Keywords:
eodysone-inducible expression system; inducible expression; serotonin receptor; tetracycline-inducible expression system;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
46
Recensione:
Indirizzi per estratti:
Indirizzo: Haegeman, G State Univ Ghent, Dept Mol Biol, KL Ledeganckstr 35, B-9000 Ghent, Belgium State Univ Ghent KL Ledeganckstr 35 Ghent Belgium B-9000 gium
Citazione:
K. Van Craenenbroeck et al., "Evaluation of the tetracycline- and ecdysone-inducible systems for expression of neurotransmitter receptors in mammalian cells", EUR J NEURO, 14(6), 2001, pp. 968-976

Abstract

Establishing a stable cell line that expresses a particular protein of interest is often a laborious and time-consuming experience. With constitutiveexpression systems, a gradual loss of the highly expressing clones over a given time span and/or a severe counter-selection due to toxicity of the expressed protein for the host cell line are major drawbacks. In both cases, inducible expression systems offer a valuable alternative. Over the years, many regulated expression systems have been developed and evaluated. In thepresent study, we compare the efficiency, the advantages and the drawbacksof a tetracycline- and an ecdysone-inducible system for expression of the reporter protein chloramphenicol acetyltransferase and of different G-protein-coupled serotonin (5-HT) receptors. A high level of expression of different 5-HT receptors was obtained with the tetracycline-inducible system. In the cell line L929, which stably expresses the tetracycline-responsive transactivator, a maximum ligand binding of 20 000 and 9500 fmol/mg protein wasmeasured for the h5-HT1B and h5-ht(1F) receptors, respectively. In the HEK293rtTA cell line, levels of 15 700, 3000, and 9100 fmol bound ligand/mg protein were obtained for the h5-HT1B, h5-ht(1F) and h5-HT4b receptors, respectively. These high expression levels remained stable for several months ofcontinuous culture. Although. the ecdysone-inducible expression system wasuseful for tightly regulated expression, the levels were far lower than those obtained with the tetracycline system (e.g. 640 fmol bound ligand/mg protein for the h5-htlF receptor in HEK293EcR).

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 02/04/20 alle ore 11:35:49