SSSDScientific and Social Sustainable Development3066-89643066-8980Art and Technology10.61369/SSSD.2025150037Article一种快速构建巴斯德毕赤酵母外源基因表达载体的新方法https://artdesignp.com/journal/SSSD/1/15/10.61369/SSSD.2025150037许岚,余明华,钟星20251152025-10-14巴斯德毕赤酵母（Pichia pastoris）表达系统是目前工业发酵大规模生产重组蛋白最具吸引力的真核表达系统之一。本研究利用两步PCR 方法快速构建了毕赤酵母线性表达载体，并以黑曲霉（Aspergillus niger）葡萄糖氧化酶AGOX 为例，成功构建了AGOX 毕赤酵母线性表达载体，实现了其在Pichia pastorisGS115中的高效表达。该方法较传统酶切- 连接克隆方法简单、快速，为本实验室所首次报导，该技术为高通量大规模毕赤酵母表达重组蛋白奠定了基础。毕赤酵母,重叠PCR,重组蛋白药物,载体

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