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    • 2. 发明申请
    • SHORTENED PHOSPHOGLYCERATE KINASE PROMOTER
    • 漂白的磷酸酯激酶促进剂
    • WO1987003008A1
    • 1987-05-21
    • PCT/US1986002443
    • 1986-11-13
    • INTERNATIONAL GENETIC ENGINEERING, INC.LEE, Jar-HowBLAIR, LindleyHORWITZ, ArnoldKODURI, Raju, Kanaka
    • INTERNATIONAL GENETIC ENGINEERING, INC.
    • C12P21/00
    • C12N15/81C07K14/43C07K2319/00C07K2319/02C07K2319/035C07K2319/036C12N15/69
    • A shortened PGK promoter of the PGK gene comprising less than 500 base pairs of the proximal end of the 5' region of the PGK gene. The invention also provides for vehicles capable of replication and expression comprising said shortened PGK promoter operably linked to a structural gene and operably linked to a secretion signal linked to a structural gene, and also provides for hosts transformed with the vehicle. In one embodiment of the invention disclosed, the shortened PGK promoter is used with a process for regulating yeast plasmid copy number by two selectable markers, the first marker having normal or high level expression from its promoter, the second marker, leu2-d, having low level expression from its promoter. Expression of a structural gene on this plasmid system can be increased by lowering the level of the second metabolite (leucine) in the growth medium, thereby selecting for an increased copy number of the second selectable marker (leu2-d), and therefore increased plasmid copy number, which in turn increases the number of copies of the structural gene being expressed.
    • PGK基因缩短的PGK启动子包含少于500个碱基对的PGK基因的5'区域的近端。 本发明还提供能够复制和表达的载体,其包含与结构基因可操作地连接的所述缩短的PGK启动子,并可操作地连接到与结构基因连锁的分泌信号,并且还提供用载体转化的宿主。 在所公开的本发明的一个实施方案中,缩短的PGK启动子与通过两个可选择标记物调节酵母质粒拷贝数的方法一起使用,第一标记物具有来自其启动子的正常或高水平表达,第二标记leu2-d具有 来自其启动子的低水平表达。 可以通过降低生长培养基中第二代谢物(亮氨酸)的水平来提高该质粒系统上的结构基因的表达,从而选择增加的第二选择标记(leu2-d)的拷贝数,因此增加质粒 拷贝数,这又增加了正在表达的结构基因的拷贝数。