酿酒酵母亮氨酸2基因与大肠杆菌β-半乳糖苷酶基因的融合。
Fusion of the Saccharomyces cerevisiae leu2 gene to an Escherichia coli beta-galactosidase gene.
作者信息
Martinez-Arias A E, Casadaban M J
出版信息
Mol Cell Biol. 1983 Apr;3(4):580-6. doi: 10.1128/mcb.3.4.580-586.1983.
Abstract
The promoter and translation initiation region of the Saccharomyces cerevisiae leu2 gene was fused to the Escherichia coli beta-galactosidase gene. This fusion located the control region of the leu gene and orientated its direction of expression. When the fusion was placed into yeast cells, beta-galactosidase was expressed under the same regulatory pattern as the original leu2 gene product: its synthesis was repressed in the presence of leucine and threonine. Sensitive chromogenic substrates for beta-galactosidase were used to detect expression in isolated colonies growing on agar medium. Mutant yeast cells with increased beta-galactosidase activity were identified by the color of the colonies they formed. One class of mutants obtained appeared to affect ars1 plasmid maintenance, and another class appeared to affect beta-galactoside uptake.
摘要
将酿酒酵母亮氨酸2基因的启动子和翻译起始区域与大肠杆菌β-半乳糖苷酶基因融合。这种融合确定了亮氨酸基因的控制区域并确定了其表达方向。当将该融合体导入酵母细胞时，β-半乳糖苷酶以与原始亮氨酸2基因产物相同的调控模式表达：在亮氨酸和苏氨酸存在的情况下其合成受到抑制。使用对β-半乳糖苷酶敏感的显色底物来检测在琼脂培养基上生长的分离菌落中的表达。通过它们形成的菌落颜色鉴定出具有增加的β-半乳糖苷酶活性的突变酵母细胞。获得的一类突变体似乎影响ars1质粒的维持，另一类似乎影响β-半乳糖苷的摄取。
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本文引用的文献
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Transposed LEU2 gene of Saccharomyces cerevisiae is regulated normally.酿酒酵母的转座LEU2基因调控正常。
J Bacteriol. 1980 Nov;144(2):852-5. doi: 10.1128/jb.144.2.852-855.1980.
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Analysis of gene control signals by DNA fusion and cloning in Escherichia coli.通过在大肠杆菌中进行DNA融合和克隆分析基因控制信号。
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Regulation of HIS4-lacZ fusions in Saccharomyces cerevisiae.酿酒酵母中HIS4-lacZ融合基因的调控
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Overproduction and control of the LEU2 gene product, beta-isopropylmalate dehydrogenase, in transformed yeast strains.转化酵母菌株中LEU2基因产物β-异丙基苹果酸脱氢酶的过量生产与调控
J Biol Chem. 1982 Jan 10;257(1):39-41.
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Yeast genes fused to beta-galactosidase in Escherichia coli can be expressed normally in yeast.在大肠杆菌中与β-半乳糖苷酶融合的酵母基因能够在酵母中正常表达。
Proc Natl Acad Sci U S A. 1981 Apr;78(4):2460-4. doi: 10.1073/pnas.78.4.2460.
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