Addition of Signal Leader Sequences to the N-Termini of Olfactory Receptor Proteins Enhances Their Expression in Xenopus Oocyte
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概要
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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 Vcorresponded to the signal leader (SL) sequences of 5HT_3R 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, only ODR-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.
- 社団法人日本農芸化学会の論文
- 2000-08-23
著者
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Abe K
Department Of Applied Biological Chemistry Graduate School Of Agricultural And Life Sciences The Uni
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ABE Keiko
Department of Pathology, Tohoku University Graduate School of Medicine
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Abe K
Univ. Tokyo Tokyo Jpn
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Abe Keiko
Agricultural And Life Sciences Dep. Of Applied Biological Chemistry
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EMORI Yasufumi
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The U
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Abe K
Univ Tokyo Tokyo Jpn
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Abe Keiko
Department Of Cardiovascular Physiology And Medicine Graduate School Of Biomedical Sciences Hiroshim
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YASUOKA Akihito
Bio-oriented Technology Research Advancement Institution
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Abe K
Faculty Of Pharmacy Musashino University
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Emori Yasufumi
Department Of Applied Biological Chemistry Graduate School Of Agricultural And Life Sciences The Uni
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Yasuoka Akihito
Bio-oriented Technology Research Advancement Institution The University Of Tokyo
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Abe Keiko
Department Of Agricultural Chemistry The University Of Tokyo
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