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Titolo:
Addition of signal leader sequences to the N-termini of olfactory receptorproteins enhances their expression in Xenopus oocyte
Autore:
Yasuoka, A; Emori, Y; Abe, K;
Indirizzi:
Univ Tokyo, Grad Sch Agr & Life Sci, Dept Appl Biol Chem, Bunkyo Ku, Tokyo1138657, Japan Univ Tokyo Tokyo Japan 1138657 Biol Chem, Bunkyo Ku, Tokyo1138657, Japan Biooriented Technol Res Advancement Inst, Oomiya, Saitama 3318537, Japan Biooriented Technol Res Advancement Inst Oomiya Saitama Japan 3318537 pan Univ Tokyo, Fac Sci, Dept Biophys & Biochem, Bunkyo Ku, Tokyo 1130033, Japan Univ Tokyo Tokyo Japan 1130033 Biochem, Bunkyo Ku, Tokyo 1130033, Japan
Titolo Testata:
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
fascicolo: 8, volume: 64, anno: 2000,
pagine: 1688 - 1695
SICI:
0916-8451(200008)64:8<1688:AOSLST>2.0.ZU;2-2
Fonte:
ISI
Lingua:
ENG
Soggetto:
FUNCTIONAL EXPRESSION; ODORANT RECEPTOR; ENDOPLASMIC-RETICULUM; MOLECULAR-CLONING; ORYZIAS-LATIPES; DANIO-RERIO; MEDAKA FISH; MEMBRANE; CELLS; IDENTIFICATION;
Keywords:
olfactory receptor; signal leader sequence; heterologous expression; Xenopus oocyte;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
36
Recensione:
Indirizzi per estratti:
Indirizzo: Yasuoka, A Univ Tokyo, Grad Sch Agr & Life Sci, Dept Appl Biol Chem, Bunkyo Ku, 1-1-1Yayoi, Tokyo 1138657, Japan Univ Tokyo 1-1-1 Yayoi Tokyo Japan 1138657 okyo 1138657, Japan
Citazione:
A. Yasuoka et al., "Addition of signal leader sequences to the N-termini of olfactory receptorproteins enhances their expression in Xenopus oocyte", BIOS BIOT B, 64(8), 2000, pp. 1688-1695

Abstract

Several recent papers have reported the difficulties in expressing olfactory receptor proteins (ORs) in heterologous systems, and proposed that some sequences in ORs have negative effects on their efficient expression, To obtain an efficient expression system of ORs, we modified N-terminal sequences of ORs through the addition of exogenous sequences. Three kinds of sequences, designated as 5HT, V, and VL, were used. 5HT and V corresponded to thesignal leader (SL) sequences of 5HT R-3 and VIPR, respectively. VL corresponded to the first extracellular region of VIPR containing the SL sequence and three potential asparagine- (Asn-) linked glycosylation sites. The myc epitope was also added to the C-termini of the sequences. Several ORs including 17 of rat, GUST43 of rat, Y1 of medaka, FOR1-3 of pufferfish, 47E of carp, and ODR-10 of nematode were subjected to the modifications, and the RNAs encoding modified ORs were injected into Xenopus oocytes. The membrane fraction of the oocytes were analyszed by Western blotting to examine the expression of the proteins. In the cases of ORs modified with 5HT and V, onlyODR-10 and 47E, both of which have more than two Asn-linked glycosylation sites in their extracellular regions, were detected as the bands of predicted molecular weights. On the other hand, most of the ORs modified with VL showed the bands of predicted molecular weights. These results suggest that SL sequences together with potential Asn-linked glycosylation sites have positive effects on the expression of ORs in heterologous systems.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 11/07/20 alle ore 20:34:39