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1. 发明授权

US06326164B1 Methods for determining deoxyxylulose 5-phosphate synthase activity 失效
标题翻译：测定脱氧纤维素5-磷酸合成酶活性的方法
公开(公告)号：US06326164B1
公开(公告)日：2001-12-04
申请号：US09626589
申请日：2000-07-27
申请人： John W. Rice , Andreas S. Kloti , John M. Crawford, Jr. , Beth Lanning , Sandy J. Stewart
发明人： John W. Rice , Andreas S. Kloti , John M. Crawford, Jr. , Beth Lanning , Sandy J. Stewart
IPC分类号： C12Q126
CPC分类号： C12Q1/25
摘要： The invention is directed to methods and compositions for the determination of deoxyxylulose 5-phosphate synthase (DXPS) activity. The methods and compositions of the invention are amenable to high-throughput screening assays for the identification of inhibitors and enhancers of DXPS activity. Such compounds have use in the modulation of plant and microbial growth. The compositions of the invention are DXPS fragments and chimeric polypeptides that have increased solubility as compared to the wild-type DXPS polypeptide. These DXPS fragments and chimeric polypeptides can be recombinantly expressed and purified in quantities suitable for high-throughput screening assays. The assays of the invention are based on the detection of substrates of DXPS that remain after a DXPS reaction.
摘要翻译：本发明涉及用于测定脱氧纤维素-5-磷酸合成酶（DXPS）活性的方法和组合物。本发明的方法和组合物适用于鉴定DXPS活性的抑制剂和增强剂的高通量筛选分析。这些化合物可用于植物和微生物生长的调节。本发明的组合物是与野生型DXPS多肽相比具有增加的溶解度的DXPS片段和嵌合多肽。这些DXPS片段和嵌合多肽可以以适于高通量筛选测定的量重组表达和纯化。本发明的测定是基于在DXPS反应之后残留的DXPS底物的检测。

2. 发明授权

US06605459B2 Methods for measuring cysteine and determining cysteine synthase activity 失效
标题翻译：测量半胱氨酸和测定半胱氨酸合成酶活性的方法
公开(公告)号：US06605459B2
公开(公告)日：2003-08-12
申请号：US09905290
申请日：2001-07-13
申请人： John Rice , Beth Lanning , John Crawford , Gordon Nye
发明人： John Rice , Beth Lanning , John Crawford , Gordon Nye
IPC分类号： C12N988
CPC分类号： C12N9/88 , C12Q1/48 , G01N2333/91171 , G01N2500/20
摘要： Assays, preferably high throughput assays, for determining cysteine concentration, cysteine synthase activity, and identifying herbicides, fungicides, bactericides, and insecticides. Cysteine concentration is quantitated by contacting cysteine with a coumarin dye capable of conjugating with cysteine but not to O-acetyl serine or sulfide; exciting the conjugate with UV light; and detecting fluorescent light emitted by the conjugate. Cysteine synthase activity is determined by combining O-acetyl-L-serine, sulfide and cysteine synthase to form a reaction mixture under conditions suitable for cysteine production; contacting the reaction mixture with an appropriate coumarin dye; subjecting the reaction mixture to UV light; and detecting fluorescent light emission.
摘要翻译：用于确定半胱氨酸浓度，半胱氨酸合成酶活性和鉴定除草剂，杀真菌剂，杀菌剂和杀虫剂的测定，优选高通量测定。通过使半胱氨酸与能够与半胱氨酸缀合但不与O-乙酰丝氨酸或硫化物偶联的香豆素染料接触来定量半胱氨酸浓度; 用紫外线激发共轭物; 并检测共轭物发出的荧光。通过将O-乙酰基-L-丝氨酸，硫化物和半胱氨酸合成酶组合形成半胱氨酸合成酶活性，在适合于半胱氨酸生产的条件下形成反应混合物; 使反应混合物与合适的香豆素染料接触; 使反应混合物经受紫外光; 并检测荧光发光。

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