【摘要】 为了构建巴斯德毕赤酵母通用表面展示载体,本研究利用DNA重组技术从酿酒酵母EBY100中克隆了与酵母细胞壁共价连接的甘露糖蛋白PIR的N端重复序列(pir-N),体外连接到毕赤酵母分泌表达载体pPIC9K中,得到新型巴斯德毕赤酵母表面展示的通用载体pPIC9K-Pir-N,并将β-葡萄糖苷酸酶基因与pir-N端融合。结果表明β-葡萄糖苷酸酶已成功展示到巴斯德毕赤酵母细胞表面,使巴斯德毕赤酵母细胞具有了β-葡萄糖苷酸酶催化活性,研究成果为其他生物催化剂的酵母表面展示提供技术平台。更多还原

【Abstract】 To construct the general vector of Pichia pastoris cell surface display,in this study,the N-terminal sequence(pir-N) covalently linked to cell wall protein of Saccharomyces cerevisiae EBY100 was cloned by DNA recombination method.Then the pir-N gene was inserted into pPIC9K vector to obtain plasmid pPIC9K-Pir-N,and gus gene was fused with pir-N gene.The result suggested that GUS protein was displayed on Pichia pastoris cell surface successfully,which may provide a technological platform for the yeast cell-surface display of other biocatalyst.更多还原