- 询价
- ZYMO RESEARCH
- F9003
- 美国
- 2026年01月21日
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- 详细信息
- 文献和实验
- 技术资料
- 供应商：
简石生物
- 库存：
大量
- 英文名：
5-Fluoroorotic Acid (5-FOA)
HIGHLIGHTS
- Yeast Genetic Counter-Selection Agent: Commonly used for curing yeast strains of plasmids, plasmid shuffling, allelic replacement, and two-hybrid screens.
- Convenient: Available as a pure powder or ready-to-use solution in DMSO.
- Ultra-Pure: Determined > 98% by thin-layer chromatography (TLC), melting point, and lot comparison.
DESCRIPTION
Using 5-Fluoroorotic Acid (5-FOA) for the counter-selection of yeast is a common genetic screening method. Curing yeast strains of plasmids, plasmid shuffling, allelic replacement, and two-hybrid screens are methods that can employ the use of 5-FOA. Otherwise nontoxic to yeast, 5-FOA is converted to the toxic form (i.e., 5-flurouracil) in strains expressing the functional URA3 gene coding for orotine-5-monophosphate decarboxylase that is involved in the synthesis of uracil. Yeast strains that are phenotypically Ura+ become Ura- and 5-FOA(R) (resistant) after selection. The question of 5-FOA solubility is often raised by customers using ultra-pure (< 98%) 5-FOA powder because of its insolubility in water. Thus, we provide a 100X concentrated (100 mg/ml) 5-FOA solution in DMSO and synthetic 5-FOA monohydrate powder. Both have been tested and validated on the basis of counter selection activity.
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文献和实验激活域（AD）表达为融合蛋白。当两个蛋白相互作用时，AD和DBD间距离被拉近，从而激活报告基因的表达。 相互作用子蛋白的鉴定 用于ProQuest™系统的MaV203酵母宿主菌株含有报告基因，确保正确作用子蛋白的鉴定。酵母的基因组上整合有HIS3，VRA3和LacZ基因各自的单个拷贝。阳性作用子蛋白通过转化子在组氨酸和尿苷酸缺失培养基上的生长及X-gal对LacZ基因的诱导来鉴定。对URA3的诱导导致5-fluoroorotic acid（5FOA）转化为一种毒性化合
监测蛋白间的相互作用。例如酵母URA3基因表达产物是尿嘧啶合成所必需的， 同时它又可催化5-氟乳清酸(5-FOA)转化为有毒物质。Vidal等构建了一酵母细胞株， 其URA3的表达由含GAL4结合位点的启动子严密控制。此细胞株在缺乏尿嘧啶的培养基中培育需要GAL4激活结构域(GAD)和GAL4 DNA结合结构域(GBD)的融合蛋白的相互作用的表达。而在含5-FOA的完全培养基中则受GAD和GBD融合蛋白相互作用的抑制。因此可通过筛选5-FOA抗性克隆从随机突变库中鉴定阻断蛋白相互作用的突变体。
5-FOA plates (0.5 liter): Agar: 15 g Bacto-agar 300 ml H20 in 500 ml flask Autoclave 25 min. FOA mix: 0.85 g Yeast Nitrogen base - amm. sulfate and amino acids 2.5 g Ammonium sulfate 0.3 g amino acid mix (from above) 25 ml
