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    • 7. 发明授权
    • Composite yeast vectors
    • 复合酵母载体
    • US5013652A
    • 1991-05-07
    • US918147
    • 1986-10-14
    • Robert L. StrausbergSusan L. Strausberg
    • Robert L. StrausbergSusan L. Strausberg
    • C07K14/39C07K14/81C12N15/62C12N15/81
    • C07K14/8128C07K14/39C12N15/625C12N15/81C07K2319/036C07K2319/81
    • A DNA sequence comprising:(a) a hybrid first yeast promoter comprising the functional segment of the DNA sequence encoding the upstream activation site of a gene selected from the group consisting of GAL1, GAL7 and GAL10, operably linked to the functional segment of the transcription initiation site of MF-.alpha.-1;(b) the functional segment of a signal sequence substantially comprising preferred yeast codons;(c) the functional segment of the DNA sequence comprising a Saccharomyces GAPDH transcription terminator;(d) the functional segment of the Saccharomyces 2.mu. circle replication origin; and(e) the functional segment of the yeast LEU2-d selectable marker. The DNA sequence may further comprise, inserted in phase between the signal sequence and transcription terminator, the functional segment of a DNA sequence encoding a heterologous target peptide. DNA expression vectors comprising these sequences, DNA expression systems comprising these vectors and further comprising a source of the peptide encoded by the DNA sequence of the GAL4 structural gene, yeast hosts transformed with said expression vectors or expression systems, methods of producing heterologous target peptides in yeast hosts employing the sequences, vectors and systems of the invention, and heterologous target peptides produced according to these methods are disclosed. By means of the present invention, ultrahigh (greater than 1 gram/liter, or at least 10% of total yeast cell protein) expression of heterologous peptides may be effected.
    • 一种DNA序列,其包含:(a)混合的第一酵母启动子,其包含编码选自GAL1,GAL7和GAL10的基因的上游激活位点的DNA序列的功能区段,其可操作地连接到转录的功能区段 MF-α-1的起始位点; (b)基本上包含优选的酵母密码子的信号序列的功能片段; (c)包含酵母GAPDH转录终止子的DNA序列的功能区段; (d)酵母菌2亩复制起源的功能区段; 和(e)酵母LEU2-d选择标记的功能区段。 DNA序列可以进一步包含在信号序列和转录终止子之间的相位插入编码异源靶肽的DNA序列的功能区段。 包含这些序列的DNA表达载体,包含这些载体的DNA表达系统,并且还包含由所述GAL4结构基因的DNA序列编码的肽的来源,用所述表达载体或表达系统转化的酵母宿主,在所述DNA表达系统中产生异源靶肽的方法 公开了使用本发明的序列,载体和系统的酵母宿主,以及根据这些方法生产的异源靶肽。 通过本发明,可以实现异源肽的超高(大于1克/升,或总酵母细胞蛋白的至少10%)的表达。