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19 条记录 (耗时 0.03 秒)

    Nucleic acid detection
    3.
    发明授权
    Nucleic acid detection 失效
    核酸检测

    公开(公告)号：US06335162B1

    公开(公告)日：2002-01-01

    申请号：US09042287

    申请日：1998-03-13

    申请人： John W. Shultz ,  Michelle A. Nelson ,  Donna M. Leippe ,  Martin K. Lewis ,  Lisa S. Nelson

    发明人： John W. Shultz ,  Michelle A. Nelson ,  Donna M. Leippe ,  Martin K. Lewis ,  Lisa S. Nelson

    IPC分类号： C12Q168

    CPC分类号： C12Q1/6823 ,  C12Q1/04 ,  C12Q1/66 ,  C12Q1/68 ,  C12Q1/6827 ,  Y10S435/81 ,  C12Q2535/131 ,  C12Q2521/319 ,  C12Q2535/107

    摘要： This invention discloses methods, compositions and kits for the detection of extremely low levels of nucleic acid, cells and cellular material in biological samples. The nucleic acid detection systems utilize either the pyrophosphorolysis reaction catalyzed by various polymerases or nuclease digestion coupled with pyrophosphorylation catalyzed by phosphoribosylpyrophosphate synthetase to produce either deoxyribonucleoside triphosphates or ribonucleoside triphosphates. dNTPs are transformed to ATP by the action of nucleoside diphosphate kinase. The ATP produced by these reactions may be detected by luciferase or NADH based detection systems. If more sensitive detection is required, schemes for the amplification of NTPs and dNTPS are provided. A detection system for cells or cellular material in a sample is provided wherein AMP and a high energy phosphate donor added to a sample are converted to ATP by the action of endogenous enzymes, followed by detection of the ATP.

    摘要翻译： 本发明公开了用于检测生物样品中极低水平的核酸，细胞和细胞物质的方法，组合物和试剂盒。 核酸检测系统利用通过各种聚合酶催化的焦磷酸裂解反应或核酸酶消化与磷酸核糖焦磷酸合成酶催化的焦磷酸化反应产生脱氧核糖核苷三磷酸或核糖核苷三磷酸。 dNTPs通过核苷二磷酸激酶的作用转化为ATP。 由这些反应产生的ATP可以通过荧光素酶或基于NADH的检测系统来检测。 如果需要更灵敏的检测，则提供扩增NTPs和dNTPS的方案。 提供样品中的细胞或细胞材料的检测系统，其中通过内源酶的作用将AMP和加入样品的高能量磷酸盐供体转化为ATP，随后检测ATP。

    Polymerase stabilization by polyethoxylated amine surfactants
    4.
    发明授权
    Polymerase stabilization by polyethoxylated amine surfactants 有权
    通过聚乙氧基化胺表面活性剂稳定聚合酶

    公开(公告)号：US06242235B1

    公开(公告)日：2001-06-05

    申请号：US09338174

    申请日：1999-06-22

    申请人： John W. Shultz ,  Fen Huang

    发明人： John W. Shultz ,  Fen Huang

    IPC分类号： C12N912

    CPC分类号： C12N9/12 ,  C12N9/1241 ,  C12N9/1252 ,  C12N9/1276 ,  C12N9/96

    摘要： The present invention provides methods and compositions for protein stabilization, particularly the stabilization of polymerases in aqueous solutions with cationic surfactants. The present invention further provides cationic surfactants, including polyethoxylated amines, that stabilize thermostable and thermolabile enzymes in solution. These surfactants stabilize the activity of various enzymes, including thermostable DNA polymerases, thermolabile DNA polymerases and reverse transcriptases.

    摘要翻译： 本发明提供用于蛋白质稳定化的方法和组合物，特别是在具有阳离子表面活性剂的水溶液中稳定聚合酶。 本发明还提供了在溶液中稳定热稳定和不耐热酶的阳离子表面活性剂，包括聚乙氧基化胺。 这些表面活性剂稳定各种酶的活性，包括热稳定的DNA聚合酶，不耐热的DNA聚合酶和逆转录酶。

    Proteases of altered stability to autolytic degradation
    5.
    发明授权
    Proteases of altered stability to autolytic degradation 失效
    自由降解稳定性变化的蛋白酶

    公开(公告)号：US5543302A

    公开(公告)日：1996-08-06

    申请号：US159211

    申请日：1993-11-30

    申请人： George Boguslawski ,  Josephine Grosch ,  John W. Shultz ,  Sethuraman Subramanian

    发明人： George Boguslawski ,  Josephine Grosch ,  John W. Shultz ,  Sethuraman Subramanian

    IPC分类号： C11D3/386 ,  C12N9/50 ,  C12N9/54 ,  C12N9/56 ,  C12P21/00 ,  C07K14/00 ,  C12N9/00

    CPC分类号： C12N9/54 ,  C11D3/38618 ,  C12N9/50

    摘要： Proteases are susceptible to autolytic degradation. The method of the invention provides a method to identify susceptible autolysis sites which are cleaved to produce a few discrete fragments. Once identified, amino acid(s) at or in the vicinity of the cleavage site may be chemically modified or the DNA sequence capable of encoding the amino acid present at the site may be changed or deleted by site directed mutagenesis to produce an autolysis stable protease. The invention is particularly applicable to alkaline proteases which are produced in large quantities and used as detergent additives. Stable alkaline proteases could then be used in liquid detergent formulations.

    摘要翻译： 蛋白酶易于自溶降解。 本发明的方法提供了一种识别敏感的自溶位点的方法，其被切割以产生几个离散的片段。 一旦鉴定，可以化学修饰在切割位点处或其附近的氨基酸，或者可以通过定点诱变来改变或删除存在于位点的氨基酸的DNA序列以产生自溶稳定蛋白酶 。 本发明特别适用于大量生产并用作洗涤剂添加剂的碱性蛋白酶。 稳定的碱性蛋白酶可用于液体洗涤剂配方。