Expression and Purification of Cytokine Receptor Homology Domain of Human Granulocyte-Colony Stimulating Factor Receptor in Escherichia coli
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概要
- 論文の詳細を見る
In an attempt to generate a stable non-glycosylated cytokine receptor homology (CRH) domain (Tyr_<97>-Ala_<309>) of human granulocyte-colony stimulating factor (G-CSF) receptor, two free cysteines in the CRH domain were converted to serine by site-directed mutagenesis. Taking advantage of the tight regulation for the expression of T7 RNA polymerase, the mutated CRH domain was successfully expressed in Escherichia coli (E. coli) with a pelB signal sequence at its NH_2-terminus and with a His tag at its COOH-terminus. The processed and secreted CRH domain after solubilization and in vitro refolding retained G-CSF binding activity, and its yield (〜40μg/30 ml culture) was more than 100-fold higher than that of the mouse CRH domain expressed by the MalE fusion system in E. coli.
- 社団法人日本農芸化学会の論文
- 1998-09-23
著者
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Tanaka Ryoichi
Faculty Of Agriculture Kagoshima University
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Takagi H
Department Of Food Science And Technology Faculty Of Agriculture Nagoya University:(present Address)
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TOKUNAGA Masao
Laboratory of Applied and Molecular Microbiology, Faculty of Agriculture, Kagoshima University
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TOKUNAGA Hiroko
Laboratory of Applied and Molecular Microbiology, Faculty of Agriculture, Kagoshima University
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TANAKA Ryoichi
Laboratory of Applied and Molecular Microbiology, Faculty of Agriculture, Kagoshima University
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HARA Shinichi
Amgen Inc., Amgen Center
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ARAKAWA Tsutomu
Amgen Inc., Amgen Center
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Tokunaga M
Applied And Molecular Microbiology Faculty Of Agriculture Kagoshima University
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Tokunaga Masao
Laboratories Of Molecular Genetics Mitsubishi Kasei Institute Of Life Sciences
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Tanaka Ryutaro
Faculty Of Agriculture Kagoshima University
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Arakawa T
Alliance Protein Laboratories
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Arakawa Tsutomu
Amgen Inc.
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Hara Shinichi
Amgen Inc. Amgen Center
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Tokunaga Hiroko
Laboratory Of Applied And Molecular Microbiology Faculty Of Agriculture Kagoshima University
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Arakawa T
Allaiance Protein Laboratories
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Tokunaga Masao
Faculty Of Agri. Kagoshima Univ.
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