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    • 9. 发明申请
    • METHODS FOR INCREASING EXPRESSION OF GENES IN A FUNGAL CELL
    • 提高真核细胞中基因表达的方法
    • WO2008008967A2
    • 2008-01-17
    • PCT/US2007073486
    • 2007-07-13
    • NOVOZYMES INCNOVOZYMES ASYAVER DEBBIECHERRY BARBARABJOERNVAD MADS ESKELUND
    • YAVER DEBBIECHERRY BARBARABJOERNVAD MADS ESKELUND
    • C12N15/81C07K14/395
    • The present invention relates to methods for producing a polypeptide, comprising: (a) cultivating a fungal host cell in a medium conducive for the production of the polypeptide, wherein the fungal host cell comprises a first polynucleotide comprising a nucleic acid sequence encoding the polypeptide operably linked to a copper-inducible promoter sequence comprising a copper-responsive upstream activation sequence activated by a copper-dependent trans-acting transcription factor and a second polynucleotide comprising one or more (several) additional copper-responsive upstream activation sequences operably linked upstream to the promoter sequence, wherein the promoter sequence is foreign to the nucleic acid sequence encoding the polypeptide and the copper-responsive upstream activation sequences are responsible for copper-induced transcription of the promoter sequence, and a third polynucleotide comprising at least one copy of a gene encoding the copper-dependent trans-acting transcription factor; and (b) isolating the polypeptide from the cultivation medium.
    • 本发明涉及用于产生多肽的方法,其包括:(a)在有助于产生所述多肽的培养基中培养真菌宿主细胞,其中所述真菌宿主细胞包含第一多核苷酸,所述第一多核苷酸包含操作性编码所述多肽的核酸序列 与铜诱导型启动子序列连接,所述启动子序列包含铜依赖性反式作用转录因子激活的铜响应上游激活序列和第二多核苷酸,所述第二多核苷酸包含一个或多个(几个)额外的铜响应上游激活序列, 启动子序列,其中所述启动子序列对编码所述多肽的核酸序列是外源的并且所述铜响应上游激活序列负责所述启动子序列的铜诱导的转录,和第三多核苷酸,其包含编码所述多肽的基因的至少一个拷贝 铜依赖的反式转录转录 n因素; 和(b)从培养基中分离多肽。