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    • 6. 发明申请
    • YEAST BASED EXPRESSION OF PROTEASES AND METHODS OF USE
    • 基于YEAST的表达及其使用方法
    • US20100028892A1
    • 2010-02-04
    • US12504600
    • 2009-07-16
    • John C. ReedHideki HayashiMichael Cuddy
    • John C. ReedHideki HayashiMichael Cuddy
    • C12Q1/68
    • C12N15/1055C12N15/1086C12N15/81
    • This disclosure generally relates to components and methods of using a high throughput screening (HTS) systems for intracellular proteases, using Caspases as a prototype. Genetic systems are disclosed for monitoring exogenous caspase activation pathways in the yeast, Saccharomyces cerevisiae. The yeast-based cellular systems permit facile expression of proteases (e.g., caspase) and protease-activating proteins in combinations that reconstitute entire mammalian pathways in these simple eukaryotes. Among the assay methods integrated into the yeast system are cleavable reporter gene activators, in which protease-mediated cleavage activates a transcription factor. Exemplary systems rely, singly or in concert, on exogenous recombinant caspases and exogenous upstream activators of caspases to cleave a chimeric protein giving rise to a transcription factor which induces the expression of the LacZ and LEU2 genes. The activities of these genes result in colored cultures and impart the ability of the yeast to grow in leucine deficient media. The intensity of the color is measured by colorimetry and quantified with OD units. The OD units are directly proportional to the activity of the caspase in the system. The method of quantification is referred to as the “readout”.
    • 本公开通常涉及使用胱天蛋白酶作为原型的细胞内蛋白酶的高通量筛选(HTS)系统的组件和方法。 公开了用于监测酵母,酿酒酵母中的外源半胱天冬酶活化途径的遗传系统。 基于酵母的细胞系统允许在这些简单真核生物中重组整个哺乳动物途径的组合,容易地表达蛋白酶(例如胱天蛋白酶)和蛋白酶活化蛋白。 整合到酵母系统中的测定方法中是可裂解的报告基因激活因子,其中蛋白酶介导的切割激活转录因子。 示例性系统单独或一致地依赖于外源重组胱天蛋白酶和胱天蛋白酶的外源上游激活剂以切割嵌合蛋白,产生诱导LacZ和LEU2基因表达的转录因子。 这些基因的活性导致有色培养物,并赋予酵母在亮氨酸缺乏培养基中生长的能力。 通过比色法测量颜色的强度并用OD单位定量。 OD单位与系统中半胱天冬酶的活性成正比。 定量方法被称为“读出”。
    • 9. 发明授权
    • Silicon carbide semiconductor device
    • 碳化硅半导体器件
    • US08766278B2
    • 2014-07-01
    • US13565388
    • 2012-08-02
    • Hideki Hayashi
    • Hideki Hayashi
    • H01L29/15H01L21/8238
    • H01L27/088H01L21/8213
    • First, second, fourth, and fifth impurity regions have a first conductivity type, and a third impurity region has a second conductivity type. The first to third impurity regions reach a first layer having the first conductivity type. The fourth and fifth impurity regions are provided on a second layer. First to fifth electrodes are provided on the first to fifth impurity regions, respectively. Electrical connection is established between the first and fifth electrodes, and between the third and fourth electrodes. A sixth electrode is provided on a gate insulating film covering a portion between the fourth and fifth impurity regions.
    • 第一,第二,第四和第五杂质区具有第一导电类型,第三杂质区具有第二导电类型。 第一至第三杂质区域到达具有第一导电类型的第一层。 第四和第五杂质区设置在第二层上。 第一至第五电极分别设置在第一至第五杂质区上。 在第一和第五电极之间以及第三和第四电极之间建立电连接。 第六电极设置在覆盖第四和第五杂质区域之间的部分的栅极绝缘膜上。