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このアイテムの引用には次の識別子を使用してください: http://hdl.handle.net/10129/6147


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タイトル: A Comparative Study of The Internal Ribosome Entry Sitecontaining Expression System
著者: Murakami, Manabu
Murakami, Agnieszka A
Ohba, Takayoshi
著者所属: Department of Pharmacology, Hirosaki University Graduate School of Medicine
Department of Cellphysiology, Akita University School of Medicine
Department of Cellphysiology, Akita University School of Medicine
発行日: 5-Oct-2017
出版者: 弘前大学大学院医学研究科・弘前医学会
誌名: 弘前医学
巻: 68
号: 1
開始ページ: 44
終了ページ: 51
ISSN: 0439-1721
キーワード: eukaryotic expression vector
bicistronic expression vector
green fluorescent protein
internal ribosome entry site.
抄録: We analyzed transient and stable eukaryotic gene expression using internal ribosome entry site (IRES) and green fluorescent protein (GFP)-S65T (IRES-GFP) expression systems. The IRES-containing vector enables simple construction of a plasmid and easy detection of transfected cells, as well as an estimation of the levels of the gene of interest. This system is useful for measuring changes in the intracellular calcium concentration using the fluorescent calcium indicator Fura-2 AM, although subcellular localization of the protein of interest is not detectable. Importantly, the IRES-GFP plasmid showed delayed expression and an approximate 40% reduction in fluorescence intensity, which suggests that usage of IRES-containing expression system might cause under-estimation of desired gene. Although this technique has some disadvantages, the IRES-containing vector is simple and straightforward for gene expression analysis, comparing with co-expression procedure of multiple plasmids.
URI: http://hdl.handle.net/10129/6147
テキストバージョン: publisher

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