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    • 86. 发明申请
    • HOST CELLS AND METHODS OF USE
    • 宿主细胞和使用方法
    • US20120142895A1
    • 2012-06-07
    • US13203056
    • 2010-02-24
    • Yong Hwan JinJames D. JowettAlexander H. TaylorYuan Zhu
    • Yong Hwan JinJames D. JowettAlexander H. TaylorYuan Zhu
    • C07K14/00C12P21/00C12N1/19
    • C12N15/815C07K14/605C07K2319/31C12N9/1051C12N9/60
    • The present invention provides genetically modified Pichia strains wherein at least one nucleic acid sequence encoding a functional gene product and/or at least one nucleic acid sequence necessary for expression of at least one functional gene product in said Pichia strain is genetically modified, wherein said gene product is responsible for proteolysis and/or glycosylation in said genetically modified Pichia strain. In particular, Pichia strains are provided wherein nucleic acid sequence encoding a functional gene product or expression of said gene product are genetically modified: PEP4, PRB1, YPS1, YPS2, YMP1, YMP2, YMP3 and PMT4. Also provided herein are genetically modified host cells wherein wild type parent of said genetically modified host cell comprises a gene encoding a polypeptide having at least 60% sequence identity to amino acids 1-865 of SEQ ID NO:4, SEQ ID NO:6 and SEQ ID NO:8 wherein said gene is genetically modified in the genome of said host cell such that the gene product is reduced or eliminated in said genetically modified host cell compared with said wild type host cell.
    • 本发明提供了遗传修饰的毕赤酵母菌株,其中至少一个编码功能性基因产物的核酸序列和/或在所述毕赤酵母属菌株中表达至少一种功能性基因产物所必需的至少一种核酸序列被遗传修饰,其中所述基因 产物负责所述遗传修饰的毕赤酵母菌株中的蛋白水解和/或糖基化。 具体而言,提供了编码功能性基因产物的核酸序列或所述基因产物的表达被遗传修饰的毕赤酵母菌株:PEP4,PRB1,YPS1,YPS2,YMP1,YMP2,YMP3和PMT4。 本文还提供遗传修饰的宿主细胞,其中所述经遗传修饰的宿主细胞的野生型亲本包含编码与SEQ ID NO:4,SEQ ID NO:6的氨基酸1-865至少60%序列同一性的多肽的基因和 SEQ ID NO:8,其中所述基因在所述宿主细胞的基因组中被遗传修饰,使得与所述野生型宿主细胞相比,在所述遗传修饰的宿主细胞中基因产物被还原或消除。