Cloning and Expression of Bioactive Human Granulocyte Colony Stimulating Factor in Pichia pastoris
學年 99
學期 1
出版(發表)日期 2010-08-01
作品名稱 Cloning and Expression of Bioactive Human Granulocyte Colony Stimulating Factor in Pichia pastoris
作品名稱(其他語言)
著者 簡素芳; Chien, Su-Fang
單位 淡江大學化學學系
出版者 臺北市:中國化學學會
著錄名稱、卷期、頁數 Journal of the Chinese Chemical Society=中國化學會會誌 57(4B), pp.850-856
摘要 For over ten years, recombinant human Granulocyte Colony Stimulating Factor (hG-CSF) has been broadly used in clinical applications. Many sources of G-CSF have the disadvantage of being in intracellular or aggregated form. In this paper, we report the molecular cloning and gene expression in the protease resistant strain of yeast (SMD1168). More than 90% of the G-CSF protein is in secreted form and about 100 mg of the biologic active human G-CSF can be obtained from two liters of bioreactor. The recombinant G-CSF promotes HL-60 (Human promyelocytic cell) cell proliferation and elevates the mito-chondrial succinate dehydrogenase activity. The dose effects of rG-CSF on cell proliferation responses to the total numbers of the receptors on the cell surfaces have been investigated and the quantitative results were obtained. The receptors are saturated byG-CSF atthe50 ng/mLfor 1x 10^5 cells. The prompt increasing in the succinate dehydrogenas activity was also observed. This effect is comparable to that from the commercial source. We have successfully cloned and expressed a bioactive rhG-CSF from the medium in the Pichia pastoris system. This specific product is expected to meet the clinical applications in the treatments of neutropenia, acute cerebral ischemia and other neuro-degenerated diseases.
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語言 en
ISSN 0009-4536
期刊性質 國內
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國別 TWN
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出版型式 ,電子版,紙本
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