公开(公告)号：US5208332A

公开(公告)日：1993-05-04

申请号：US811217

申请日：1991-12-20

申请人： Babetta L. Marrone ,  Daniel J. Simpson ,  Clifford J. Unkefer ,  Thomas W. Whaley

发明人： Babetta L. Marrone ,  Daniel J. Simpson ,  Clifford J. Unkefer ,  Thomas W. Whaley

IPC分类号： C12Q1/60

CPC分类号： C12Q1/60

摘要： An optical probe enables the study of enzyme activity by absorbance spectroscopy or by sensitive fluorescence methods. In particular, the probe provides the ability to monitor the activity of cytochrome P-450.sub.scc enzyme, the rate limiting enzyme for steroid biosynthesis. Located on the inner mitochondrial membrane, P-450.sub.scc catalyzes the conversion of cholesterol to pregnenolone and isocapraldehyde by sequential oxidations of the cholesterol side chain. The fluorogenic probe includes a cholesterol-like steroid linked to a chromophore through a linking group. The chromophore is selected to have little optical response when linked to the steroid substrate and an enhanced optical response when cleaved from the substrate and linking group. Thus, a fluorescent anion that can be optically detected is generated by the side-chain cleavage reaction during steroidogenesis.

摘要翻译： 光学探针能够通过吸收光谱法或灵敏荧光法研究酶活性。 特别地，探针提供了监测细胞色素P-450scc酶的活性的能力，其是用于类固醇生物合成的限速酶。 位于内线粒体膜上，P-450scc通过胆固醇侧链的顺序氧化催化胆固醇转化为孕烯醇酮和异己醛。 荧光探针包括通过连接基团与发色团连接的胆固醇样类固醇。 选择发色团在与类固醇底物连接时具有很小的光学响应，并且当从底物和连接基团切割时具有增强的光学响应。 因此，可以通过在类固醇发生过程中的侧链切割反应产生可以被光学检测的荧光阴离子。

公开(公告)号：US5405747A

公开(公告)日：1995-04-11

申请号：US208506

申请日：1994-03-07

申请人： James H. Jett ,  Richard A. Keller ,  John C. Martin ,  Richard G. Posner ,  Babetta L. Marrone ,  Mark L. Hammond ,  Daniel J. Simpson

发明人： James H. Jett ,  Richard A. Keller ,  John C. Martin ,  Richard G. Posner ,  Babetta L. Marrone ,  Mark L. Hammond ,  Daniel J. Simpson

IPC分类号： C12Q1/68 ,  C12P19/34 ,  G01N33/48

CPC分类号： C12Q1/6816 ,  C12Q1/6869 ,  Y10T436/143333

摘要： Method for rapid-base sequencing in DNA and RNA with two-base labeling and employing fluorescent detection of single molecules at two wavelengths. Bases modified to accept fluorescent labels are used to replicate a single DNA or RNA strand to be sequenced. The bases are then sequentially cleaved from the replicated strand, excited with a chosen spectrum of electromagnetic radiation, and the fluorescence from individual, tagged bases detected in the order of cleavage from the strand.

摘要翻译： 在DNA和RNA中进行快速碱基测序的方法，采用双碱基标记，并采用双波长单分子荧光检测。 用于接受荧光标记的碱基用于复制要测序的单个DNA或RNA链。 然后从复制的链中顺序地切割碱基，用所选择的电磁辐射谱激发，并且以从链断裂的顺序检测来自单个的标记碱基的荧光。

公开(公告)号：US5558998A

公开(公告)日：1996-09-24

申请号：US486400

申请日：1995-06-05

申请人： Mark L. Hammond ,  James H. Jett ,  Richard A. Keller ,  Babetta L. Marrone ,  John C. Martin

发明人： Mark L. Hammond ,  James H. Jett ,  Richard A. Keller ,  Babetta L. Marrone ,  John C. Martin

IPC分类号： C12Q1/68 ,  G01N15/14 ,  G01N33/483 ,  G01N35/08

CPC分类号： C12Q1/6827 ,  G01N15/14 ,  Y10T436/117497

摘要： A method is provided for sizing DNA fragments using high speed detection systems, such as flow cytometry to determine unique characteristics of DNA pieces from a sample. In one characterization the DNA piece is fragmented at preselected sites to produce a plurality of DNA fragments. The DNA piece or the resulting DNA fragments are treated with a dye effective to stain stoichiometrically the DNA piece or the DNA fragments. The fluorescence from the dye in the stained fragments is then examined to generate an output functionally related to the number of nucleotides in each one of the DNA fragments. In one embodiment, the intensity of the fluorescence emissions from each fragment is linearly related to the fragment length. The distribution of DNA fragment sizes forms a characterization of the DNA piece for use in forensic and research applications.

摘要翻译： 提供了使用高速检测系统（例如流式细胞术）来定尺寸DNA片段的方法，以确定样品中DNA片段的独特特征。 在一个表征中，DNA片段在预选位点被片段化以产生多个DNA片段。 用有效染色DNA片段或DNA片段化学计量的染料处理DNA片段或得到的DNA片段。 然后检查染色片段中来自染料的荧光，以产生功能上与每个DNA片段中的核苷酸数相关的输出。 在一个实施方案中，来自每个片段的荧光发射强度与片段长度线性相关。 DNA片段大小的分布形成了用于法医学和研究应用的DNA片段的表征。

公开(公告)号：US5110725A

公开(公告)日：1992-05-05

申请号：US680978

申请日：1991-04-05

申请人： Babetta L. Marrone ,  Daniel J. Simpson ,  Clifford J. Unkefer ,  Thomas W. Whaley

发明人： Babetta L. Marrone ,  Daniel J. Simpson ,  Clifford J. Unkefer ,  Thomas W. Whaley

IPC分类号： C12Q1/60

CPC分类号： C12Q1/60

摘要： An optical probe enables the study of enzyme activity by absorbance spectroscopy or by sensitive fluorescence methods. In particular, the probe provides the ability to monitor the activity of cytochrome P-450.sub.scc enzyme, the rate limiting enzyme for steroid biosynthesis. Located on the inner mitochondrial membrane, P-450.sub.scc catalyzes the conversion of cholesterol to pregnenolone and isocapraldehyde by sequential oxidations of the cholesterol side chain. The fluorogenic probe includes a cholesterol-like steroid linked to a chromophore through a linking group. The chromophore is selected to have little optical response when linked to the steroid substrate and an enhanced optical response when cleaved from the substrate and linking group. Thus, a fluorescent anion that can be optically detected is generated by the side-chain cleavage reaction during steroidogenesis.

摘要翻译： 光学探针能够通过吸收光谱法或灵敏荧光法研究酶活性。 特别地，探针提供了监测细胞色素P-450scc酶的活性的能力，其是用于类固醇生物合成的限速酶。 位于内线粒体膜上，P-450scc通过胆固醇侧链的顺序氧化催化胆固醇转化为孕烯醇酮和异己醛。 荧光探针包括通过连接基团与发色团连接的胆固醇样类固醇。 选择发色团在与类固醇底物连接时具有很小的光学响应，并且当从底物和连接基团切割时具有增强的光学响应。 因此，可以通过在类固醇发生过程中的侧链切割反应产生可以被光学检测的荧光阴离子。