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題 名 | Molecular Cloning and Expression of a Fungal Immunomodulatory Protein, FIP-Fve, from Flammulina Velutipes=金針菇免疫蛋白質之分子篩選與基因表現 |
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作 者 | 柯俊良; 林頌然; 許瓊英; 高全良; 林榮耀; | 書刊名 | 臺灣醫學會雜誌 |
卷 期 | 96:7 1997.07[民86.07] |
頁 次 | 頁517-524 |
分類號 | 435.29、435.29 |
關鍵詞 | 金針菇免疫蛋白質; 分子篩選; 基因表現; Flammulina velutipes; Molecular cloning; Fungal immunomodulatory protein; |
語 文 | 英文(English) |
英文摘要 | FIP-fve, a fungal immunomodulatory protein, was isolated from the fruiting bodies of the edible mushroom, Flammulina velutipes. FIP-fve was shown to stimulate blast-forming activity of human peripheral blood lymphocytes and gene expression of interleukin-2, interferon- γ and tumor necrosis factor- α. Repeated adminstration of FIP-fve to mice inhibits the Arthur and systemic anaphylaxis reactions. FIP-fve cDNA was cloned and sequenced, and the amino acid sequence of FIP-fve deduced from the nucleotide sequence is identical to that previously determined by protein sequencing. FIP-fve cDNA was amplified by polymerase chain reaction, ligated into the expression vector, pGEX-2T, and expressed in Escherichia coli as a fusion protein of glutathione S-transferase (GST) and FIP-fve. The GST-FIP-fve fusion protein was soluble, and the yeild of recombinant FIP-fve was about 5 mg/L of induced culture. The recombinant FIP-fve was obtained by cleaving the GST-FIP-fve fusion protein with thrombin and purifing to homogeneity. The recombinant FIP-fve had about 50% of the immunomodulatory activity of the native FIP-fve. |
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