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Titolo:
Monitoring of in vitro and in vivo translation of green fluorescent protein and its fusion proteins by fluorescence correlation spectroscopy
Autore:
Nomura, Y; Tanaka, H; Poellinger, L; Higashino, F; Kinjo, M;
Indirizzi:
Hokkaido Univ, Res Inst Elect Sci, Lab Supramol Biophys, Sapporo, Hokkaido0600812, Japan Hokkaido Univ Sapporo Hokkaido Japan 0600812 poro, Hokkaido0600812, Japan Univ Tokyo, Inst Med Sci, Dept Clin Immunol, Tokyo, Japan Univ Tokyo Tokyo Japan o, Inst Med Sci, Dept Clin Immunol, Tokyo, Japan Univ Tokyo, Inst Med Sci, AIDS Res Ctr, Tokyo, Japan Univ Tokyo Tokyo Japan Tokyo, Inst Med Sci, AIDS Res Ctr, Tokyo, Japan Karolinska Inst, Med Nobel Inst, Dept Cell & Mol Biol, Stockholm, Sweden Karolinska Inst Stockholm Sweden ept Cell & Mol Biol, Stockholm, Sweden Hokkaido Univ, Sch Dent, Dept Oral Pathol, Sapporo, Hokkaido, Japan Hokkaido Univ Sapporo Hokkaido Japan al Pathol, Sapporo, Hokkaido, Japan
Titolo Testata:
CYTOMETRY
fascicolo: 1, volume: 44, anno: 2001,
pagine: 1 - 6
SICI:
0196-4763(20010501)44:1<1:MOIVAI>2.0.ZU;2-I
Fonte:
ISI
Lingua:
ENG
Soggetto:
CORRELATION MICROSCOPY; LIVING CELLS; DIFFUSION; PCR;
Keywords:
GFP; GFP-fusion protein; fluorescence correlation spectroscopy; diffusion time;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
25
Recensione:
Indirizzi per estratti:
Indirizzo: Nomura, Y Hokkaido Univ, Res Inst Elect Sci, Lab Supramol Biophys, Sapporo, Hokkaido0600812, Japan Hokkaido Univ Sapporo Hokkaido Japan 0600812 aido0600812, Japan
Citazione:
Y. Nomura et al., "Monitoring of in vitro and in vivo translation of green fluorescent protein and its fusion proteins by fluorescence correlation spectroscopy", CYTOMETRY, 44(1), 2001, pp. 1-6

Abstract

Background: Because the process of protein translation is an event of sparse molecules, the measurement requires high sensitivity. One of the candidates for studying the molecules is fluorescence correlation spectroscopy (FCS), which gleans quantitative information from fluctuating fluorescence signals in a diluted solution. Methods: Using FCS, the translation products of expression plasmid for green fluorescent protein (GFP) and its fusion proteins were measured in vitroand in vivo. Results: In in vitro translation, the number of products increased linearly for 90 min upon concentration of the plasmid. The autocorrelation function for GFP was fitted with a one-component model with a diffusion time of 0.18 ms, which was identical to the value expected from the molecular weight. In the cases of GFP- tagged hypoxia-inducible factor-la and glucocorticoidreceptor, each fitting result was significantly improved with a two-component model. The slow component with a diffusion time of G ms appeared to be related to the ribosome or polysome. In response to the addition of dexamethasone, the nuclear translocation from cytosol clearly induced the decreasein number of molecules in the focal point. Conclusions: FCS permits monitoring of the number of molecules translated in vitro and in vivo, the translation rate, and the molecular weight. Cytometry 44:1-6, 2001. (C) 2001 Wiley-Liss, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 21/09/20 alle ore 16:05:57