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Titolo:
Recombinant Sindbis virus allows expression and precise targeting of proteins of the contractile apparatus in cultured cardiomyocytes
Autore:
Datwyler, DA; Magyar, JP; Busceti, V; Hirschy, A; Perriard, JC; Bailey, JE; Eppenberger, HM;
Indirizzi:
ETH Honggerberg, Inst Cell Biol, CH-8093 Zurich, Switzerland ETH Honggerberg Zurich Switzerland CH-8093 , CH-8093 Zurich, Switzerland Swiss Fed Inst Technol, Inst Biotechnol, CH-8093 Zurich, Switzerland SwissFed Inst Technol Zurich Switzerland CH-8093 93 Zurich, Switzerland
Titolo Testata:
BASIC RESEARCH IN CARDIOLOGY
fascicolo: 6, volume: 96, anno: 2001,
pagine: 630 - 635
SICI:
0300-8428(200111)96:6<630:RSVAEA>2.0.ZU;2-V
Fonte:
ISI
Lingua:
ENG
Soggetto:
GREEN FLUORESCENT PROTEIN; ADULT-RAT CARDIOMYOCYTES; SEMLIKI-FOREST-VIRUS; GENE-EXPRESSION; HYPERTROPHIC CARDIOMYOPATHY; CARDIAC MYOCYTES; RNA REPLICONS; VECTORS; CELLS; REPLICATION;
Keywords:
cultured cardiomyocytes; Sindbis virus; gene transfer; contractile proteins; ectopic expression;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
23
Recensione:
Indirizzi per estratti:
Indirizzo: Eppenberger, HM ETH Honggerberg, Inst Cell Biol, CH-8093 Zurich, Switzerland ETH Honggerberg Zurich Switzerland CH-8093 , Switzerland
Citazione:
D.A. Datwyler et al., "Recombinant Sindbis virus allows expression and precise targeting of proteins of the contractile apparatus in cultured cardiomyocytes", BAS R CARD, 96(6), 2001, pp. 630-635

Abstract

Expression of epitope-tagged sarcomeric proteins in cardiomyocytes is a powerful approach for the characterization of interacting domains. Here, we report a new strategy for the study of the targeting of contractile proteinsin cardiomyocytes by Sindbis virus (SIN)-mediated gene transfer. Two recombinant SIN were generated, one encoding the myosin-light chain MLC3f-eGFP fusion protein (SINrep5/MLC3f-eGFP), and the other encoding the alpha -actinin-DsRed fusion protein (SINrep5/alpha -actinin-DsRed). After infection of long-term cultured neonatal and adult rat cardiomyocytes with SINrep5/MLC3f-eGFP, the exogenous MLC3f-eGFP fusion protein localized to the sarcomeres. Freshly isolated rod-shaped ventricular cardiomyocytes infected with SINrep5/alpha -actinin-DsRed exhibited a correct incorporation of the newly synthesized alpha -actinin-DsRed fusion protein at the Z-band of the sarcomere. This allows the assumption that the exogenous protein is assembled into myofibrils in living cardiomyocytes using the same molecular interactions equally to the endogenous counterpart. It has been thus demonstrated that the SIN expression system makes possible the straightforward analysis of the localization of sarcomeric proteins in cultured cardiomyocytes and may offer new possibilities for the characterization of mutant proteins involved in hypertrophic cardiomyopathies.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 04/06/20 alle ore 01:40:56