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-2 MANUFACTURING METHOD OF RECOMBINANT PROTEIN HUMAN INTERLEUKIN-2 IN PICHIA PASTORIS AND PICHIA PASTORIS EXPRESSING RECOMBINANT PROTEIN HUMAN INTERLEUKIN-2
-2 MANUFACTURING METHOD OF RECOMBINANT PROTEIN HUMAN INTERLEUKIN-2 IN PICHIA PASTORIS AND PICHIA PASTORIS EXPRESSING RECOMBINANT PROTEIN HUMAN INTERLEUKIN-2
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机译：-2重组人白细胞介素2蛋白的制备方法及表达重组人白细胞介素2蛋白的表达的毕赤酵母
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摘要
PURPOSE: A method of producing a recombinant human interleukin-2 protein is provided to effectively and economically make mass production of the recombinant interleukin-2 protein for the maintenance of physiological activation using Pichia pastoris. CONSTITUTION: A recombinant vector of pGAPZ A/rhIGF-2 contains a glyceraldehyde-3-phosphate dehydrogenase (GAPDH) promoter (GAP), an -factor leader sequence of Saccharomyces cerevisiae, a human interleukin-2 gene, and translation stop codons. A recombinant interleukin-2 protein is expressed from Pichia pastoris/rhIL-2 (KCCM10322). In a method of preparing a recombinant interleukin-2, an interleukin-2 gene is inserted into the pGAPZ A vector to make pGAPZ A/rhIL-2. The pGAPZ A/rhIL-2 is transduced into Pichia pastoris X33 to make a transformant. The transformed Pichia pastoris X33 is cultured in a medium, and Pichia pastoris/rhIL-2 with the multicopy insertion of pGAPZ A/rhIL-2 is selected. The Pichia pastoris/rhIL-2 is continuously cultured to make extracellular expressions of the interleukin-2, which are then purified.
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机译：目的：提供一种生产重组人白介素2蛋白的方法，以有效和经济地大规模生产重组白介素-2蛋白，以维持巴斯德毕赤酵母的生理活化。组成：pGAPZ A / rhIGF-2的重组载体包含3-磷酸甘油醛脱氢酶（GAPDH）启动子（GAP），酿酒酵母的因子前导序列，人白介素2基因和翻译终止密码子。从巴斯德毕赤酵母/ rhIL-2（KCCM10322）表达重组白介素-2蛋白。在制备重组白介素2的方法中，将白介素2基因插入pGAPZ A载体中以制备pGAPZ A / rhIL-2。将pGAPZ A / rhIL-2转导到巴斯德毕赤酵母X33中以形成转化体。在培养基中培养转化的巴斯德毕赤酵母X33，并选择多拷贝插入pGAPZ A / rhIL-2的巴斯德毕赤酵母/ rhIL-2。持续培养巴斯德毕赤酵母/ rhIL-2，以使白介素-2在细胞外表达，然后将其纯化。
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