【摘要】 目的:利用在巴氏毕赤酵母细胞表达重组人腺激肽释放酶2(rhK2)。方法:构建表达载体pPICZαC-mhK2及具有3个表达盒子的pPICZαC-3mhK2。采用电转法,将重组表达质粒转染毕赤酵母X33细胞,甲醇诱导表达,离子交换层析纯化rhK2蛋白;检测rhK2的酶活性。结果:获得了高表达rhK2的酵母细胞株,其酶活性接近正常精浆中hK2的活性。结论:毕赤酵母能够成功表达具有天然活性的人腺激肽释放酶2(hK2)。更多还原

【Abstract】 Objective: To investigate the expression of recombinant human glandular kallikrein 2 in the methylotropic yeast Pichia pastorsi. Methods: The expression vectors pPICZαC-mhK2 and multimers pPICZαC- 3mhK2 which have 3 expression cassettes were constructed and transferred into X33 cells by electroporation. The protein was expressed in Pichia pastorsi induced with methanol. The rhK2 was purified by anion-exchange chromatography and its activity was assayed. Result: rhK2 was obtained in Pichia pastorsi with its biological activity is almost equal to the natural hK2. Conclusion: The mature rhK2 can be expressed in Pichia pastoris.更多还原