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    • 5. 发明公开
    • Method for expression of PDGF or IGF proteins in yeast
    • Verfahren zur表达冯PDGF在Hefe中的IGR蛋白质
    • EP1365028A1
    • 2003-11-26
    • EP03012465.5
    • 1997-12-12
    • CHIRON CORPORATION
    • Tekamp-Olson, PatriciaMerryweather, James
    • C12N15/81C07K14/49C07K14/65C12N1/19
    • A61K38/1858A61K9/0014A61K9/06A61L26/00C07K14/49C07K14/65C07K2319/02C12N15/81
    • Vectors are provided for expression of heterologous mammalian proteins. Vectors of the invention comprise nucleotide sequences encoding a signal peptide sequence for a yeast secreted protein, an optional leader peptide sequence for a yeast secreted protein, a native propeptide leader sequence for a mature protein of interest, and a sequence for the mature protein of interest, all operably linked to a yeast promoter. Each of these elements is associated with a processing site recognized in vivo by a yeast proteolytic enzyme. Any or all of these processing sites may be a preferred processing site that has been modified or synthetically derived for more efficient cleavage in vivo . The vectors are useful in methods for expression of heterologous mammalian proteins and their secretion in the biologically active mature form. Particularly, vectors comprising these nucleotides coding sequences can be used to transform a yeast host cell, which can then be cultured and screened for secretion of the biologically active mature protein of interest.
    • 提供用于表达异源哺乳动物蛋白质的载体。 本发明的载体包含编码酵母分泌蛋白的信号肽序列,酵母分泌蛋白的任选的前导肽序列,感兴趣的成熟蛋白的天然前肽前导序列以及感兴趣的成熟蛋白的序列的核苷酸序列 ,全部可操作地连接到酵母启动子。 这些元素中的每一个与酵母蛋白水解酶在体内识别的处理位点相关联。 这些加工部位中的任何一个或全部可以是经修饰或合成衍生的用于在体内更有效切割的优选加工部位。 载体可用于表达异源哺乳动物蛋白质及其在生物活性成熟形式中的分泌的方法。 特别地,包含这些核苷酸编码序列的载体可用于转化酵母宿主细胞,然后可将其培养并筛选目的生物活性成熟蛋白质的分泌。