一种新型多拷贝毕赤酵母表达载体的构建

CONSTRUCTION OF A NEW MULTI-COPY PICHIA EXPRESSION VECTOR

目的:构建合适的载体用于外源基因在巴斯德毕赤酵母中的正确分泌表达。方法:利用基因重组技术,对已有载体pPIC9K进行改造:除去信号肽末端的XhoI酶切位点,在多克隆位点处添加此切点;添加一个多聚组氨酸纯化标签;重建了阅读框架。结果:成功构建了一个多拷贝毕赤酵母表达载体。结论:可以利用这个表达载体对外源基因进行分泌表达。

Objective：To construct suitable vectors for the secretory expression of heterologous proteins in Pichia pastoris. Methods：A restriction site XhoI was eliminated in restriction site the end of XhoI was secretion signal, another added in multiple cloning site;Polyhistidine 6xHis was added before multiple cloning sites and the reading frame was reconstructed. Result：A new multi-copy vector pPEX9k derived from pPIC9K was successfully constructed. Conclusion. The vector pPEX9k is suitable for the secretory expression of heterologous proteins.