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李明刚
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期刊论文
Construction of a recombinant Pichia Pastoris strain secreting insulin
，-0001，（）：
An intact insulin gene was synthesized by half-enzymic method and cloned into expression vector pPIN9K. After transferring the expression vector pPINS319 into Pichia pastoris GS115, two Pichia pastoris strains secreting high-level human insulin were obtained. The expressed human insulin proteins of recombinant 4 and 9 were 76 and 62 times higher than that of the negative control, respectively. Moreover 88%-90% of the insulin was secreted out of the cells. The actual insulin products of recombinant 4 and 9 calculated according to the sample volume reached 0.563g/l and 0.441g/l, respectively.
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