Shaw K J, Frommer B R, Anagnost J A, Narula S, Leibowitz P J
Schering Corporation, Bloomfield, NJ 07003.
DNA. 1988 Mar;7(2):117-26. doi: 10.1089/dna.1988.7.117.
Murine granulocyte-macrophage colony-stimulating factor (GM-CSF) was expressed in Saccharomyces cerevisiae using a novel regulated secretion system. This system involves the fusion of the GAL1 upstream regulatory region to the signal sequence of the alpha mating pheromone, and the integration of this GAL1:MF alpha 1 prepro:MuGM-CSF construct into the yeast chromosome. These constructs were very stable under both selective and nonselective conditions: after 30 generations of growth no plasmid loss was observed. The expression and secretion of MuGM-CSF were analyzed by biological assays and Western blots of yeast culture medium and yeast cell extracts. Expression of MuGM-CSF was regulated by galactose induction. In addition, expression levels were proportional to the number of tandem copies of the gene inserted into the yeast chromosome.
使用一种新型调控分泌系统在酿酒酵母中表达小鼠粒细胞-巨噬细胞集落刺激因子（GM-CSF）。该系统涉及将GAL1上游调控区域与α交配信息素的信号序列融合，并将此GAL1:MFα1前原：MuGM-CSF构建体整合到酵母染色体中。这些构建体在选择性和非选择性条件下都非常稳定：经过30代生长后未观察到质粒丢失。通过生物学测定以及对酵母培养基和酵母细胞提取物的蛋白质印迹分析来检测MuGM-CSF的表达和分泌。MuGM-CSF的表达受半乳糖诱导调控。此外，表达水平与插入酵母染色体中的基因串联拷贝数成正比。
