Enhanced production of anticoagulant hirudin in recombinant Saccharomyces cerevisiae by chromosomal δ-integration
Enhanced production of anticoagulant hirudin in recombinant Saccharomyces cerevisiae by chromosomal δ-integration
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DOI：
10.1016/s0168-1656(00)00376-x
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发表时间：
2001-01-23
影响因子：
4.1
通讯作者：
Seo, JH
中科院分区：
文献类型：
--
作者：
Kim, MD;Rhee, SK;Seo, JH
重组酿酒酵母菌株过量生产一种抗凝血水蛭素。酵母反转录转座子Ty1和URA3的δ -序列作为水蛭素表达盒的靶位点。使用优势选择抗生素G418成功选择高拷贝数的转化子。整合到酵母染色体f序列中的水蛭素表达盒的拷贝数在每个细胞5到10个拷贝之间。在δ整合重组酿酒葡萄球菌系统中，水蛭素的产量比基于yep的外源性水蛭素表达系统增加了两倍以上。在10份以内，水蛭素表达盒拷贝数与水蛭素表达量呈线性关系。整合到酵母染色体上的水蛭素表达带在非选择性培养条件下稳定维持。(C) 2001 Elsevier Science B.V.版权所有
Recombinant Saccharomyces cerevisiae strains were developed to overproduce an anticoagulant hirudin. The delta -sequences of the yeast retrotransposon Ty1 and URA3 were used as target sites for a hirudin expression cassette. High copy-number transformants were successfully selected using a dominant selection antibiotic, G418. The copy numbers of the hirudin expression cassette integrated into F-sequences of the yeast chromosome ranged from five to ten copies per cell. Production of hirudin in the delta -integrated recombinant S. cerevisiae system increased over two-fold compared with the YEp-based episomal hirudin expression system. A linear relationship between the copy number of the hirudin expression cassette and hirudin expression level was observed up to 10 copies. The hirudin expression cassettes integrated into the yeast chromosome were stably maintained in non-selective culture conditions. (C) 2001 Elsevier Science B.V. All rights reserved.