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11. 发明申请

US20040185443A1 Analyte-specific assays based on formation of a replicase substrate 审中-公开
标题翻译：基于复制酶底物形成的分析物特异性测定
公开(公告)号：US20040185443A1
公开(公告)日：2004-09-23
申请号：US10391368
申请日：2003-03-18
发明人： Gary A. Dahl
IPC分类号： C12Q001/68
CPC分类号： C12Q1/6816 , C12Q2531/149 , C12Q2525/161 , C12Q2521/307
摘要： Methods and compositions are provided for assaying for a target analyte in a sample by formation of a substrate for a replicase. The target analyte, if present in the sample, is first bound to a reporter probe. The reporter probe comprises a first portion and a second portion. The first portion comprises a polynucleotide that encodes at least part of a sequence for an RNA that is a substrate for replication. The second portion comprises a molecule that has affinity for an analyte. The reporter probe itself is not a substrate for a replicase. However, a replicase substrate is generated by treating reporter probe which is bound to analyte with a composition having nuclease activity in order to release the parts of the first portion, and then the released parts and one or more mononucleotides or oligonucleotides comprising missing parts of the substrate sequence are joined with a composition having ligase activity. Newly formed substrate is replicated with a replicase, and a signal is measured which indicates the presence and quantity of analyte in the sample. The invention also provides methods and compositions for simultaneously assaying for any of multiple analytes in a sample.
摘要翻译：提供方法和组合物用于通过形成用于复制酶的底物来测定样品中的目标分析物。目标分析物（如果存在于样品中）首先与报道探针结合。记录探针包括第一部分和第二部分。第一部分包含编码作为复制底物的RNA的序列的至少一部分的多核苷酸。第二部分包含对分析物具有亲和性的分子。报道探针本身不是复制酶的底物。然而，通过用与具有核酸酶活性的组合物结合分析物的报道探针来释放复制酶底物以释放第一部分的部分，然后释放的部分和一个或多个单核苷酸或寡核苷酸包含缺失的部分底物序列与具有连接酶活性的组合物连接。用复制酶复制新形成的底物，并测量表明样品中分析物的存在和数量的信号。本发明还提供用于同时测定样品中多种分析物中的任何一种的方法和组合物。

12. 发明申请

US20020192677A1 Nucleic acid amplification with DNA-dependent RNA polymerase activity of RNA replicases 失效
标题翻译：核酸扩增与RNA复制酶的DNA依赖性RNA聚合酶活性
公开(公告)号：US20020192677A1
公开(公告)日：2002-12-19
申请号：US10071057
申请日：2002-02-07
申请人： Promega Corporation
发明人： Randall L. Dimond , Steven J. Ekenberg , James R. Hartnett , Geoffrey R. Hudson , Leopoldo G. Mendoza , Katharine M. Miller , John E. Monahan , Christopher L. Jones , Mark A. Maffitt , Richard A. Martinelli , Edward E. Pahuski , James W. Schumm
IPC分类号： C12Q001/68 , C12P019/34
CPC分类号： C12Q1/6867 , C12Q1/6806 , C12Q1/6853 , C12Q2521/501 , C12Q2521/325 , C12Q2521/119 , C12Q2521/107 , C12Q2563/137 , C12Q2531/149 , C12Q2525/121
摘要： The present invention entails methods, and kits for carrying them out, based on the discovery that an RNA replicase, such as Qnull replicase, has DNA-dependent RNA polymerase (nullDDRPnull) activity with nucleic acid segments, including DNA segments and DNA:RNA chimeric segments, which comprise a 2null-deoxyribonucleotide or an analog thereof and which have sequences of RNAs that are autocatalytically replicatable by the replicase. The discovery of this DDRP activity provides methods of the invention for nucleic acid amplification wherein a nucleic acid, with a DNA segment with the sequence of an RNA that is autocatalytically replicatable by an RNA replicase, is provided as a substrate for the replicase. The replicase catalyzes synthesis, from the DNA segment, of the RNA, which the replicase then autocatalytically replicates. The invention entails use of the amplification methods in detecting nucleic acid analytes, as in nucleic acid probe hybridization assays. Such assays of the invention include those wherein a nucleic acid analyte is hybridized with one or more nucleic acid probes, which include or are processed to generate a DNA segment which is amplifiable through production from the segment, catalyzed by the DDRP activity of an RNA replicase, of an autocatalytically replicatable RNA, which is autocatalytically replicated to provide an abundance of readily detectable reporter molecules. The invention permits replacement of an RNA, that is autocatalytically replicatable with an RNA replicase and employed as a reporter or label in prior art assays, such as nucleic acid probe hybridization assays or immunoassays, with a nucleic acid comprising a DNA segment with the same base sequence as the RNA. The invention also includes the methods of the invention with Mnnull2, Conull2, or Znnull2 in the solutions in which the DDRP activity occurs.
摘要翻译：基于以下发现，本发明涉及用于携带它们的方法和试剂盒：RNA复制酶（例如Qbeta复制酶）具有核酸片段（包括DNA片段和DNA）的DNA依赖性RNA聚合酶（“DDRP”）活性的发现： RNA嵌合片段，其包含2'-脱氧核糖核苷酸或其类似物，并且具有由复制酶自动催化复制的RNA序列。该DDRP活性的发现提供了用于核酸扩增的本发明的方法，其中提供具有可由RNA复制酶自动催化复制的RNA序列的DNA区段的核酸作为复制酶的底物。复制酶催化从DNA片段合成RNA，然后复制酶自动催化复制。本发明需要使用扩增方法来检测核酸分析物，如在核酸探针杂交测定中。本发明的这种测定包括其中核酸分析物与一个或多个核酸探针杂交的那些，其包括或被加工以产生可通过片段产生扩增的DNA片段，由RNA复制酶的DDRP活性催化的自催化可复制的RNA，其被自动催化复制以提供丰富的容易检测的报道分子。本发明允许用RNA复制酶自动催化复制并在现有技术测定中用作报告物或标记的RNA，例如核酸探针杂交测定或免疫测定，其中核酸包含具有相同碱基的DNA区段序列作为RNA。本发明还包括在发生DDRP活性的溶液中具有Mn +2，Co + 2或Zn + 2的本发明的方法。

13. 发明申请

US20120322675A1 GENOME-SCALE ANALYSIS OF REPLICATION TIMING 审中-公开
标题翻译：复制时间的基因量化分析
公开(公告)号：US20120322675A1
公开(公告)日：2012-12-20
申请号：US13479686
申请日：2012-05-24
申请人： David M. Gilbert , Tyrone Ryba , Ichiro Hiratani
发明人： David M. Gilbert , Tyrone Ryba , Ichiro Hiratani
IPC分类号： C40B30/04 , G06F19/20
CPC分类号： C12Q1/6809 , C12Q1/6841 , C12Q1/6881 , C12Q2600/158 , C12Q2527/113 , C12Q2531/149 , C12Q2539/115 , C12Q2565/501
摘要： Methods for identifying and/or distinguishing a homogeneous population of cells based on their replication domain timing profile using high resolution genomic arrays or sequencing procedures are provided. These methods may be used to compare the replication timing profile for a population of cells to another replication timing profile(s), a replication timing fingerprint, and/or one or more informative segments of a replication timing fingerprint, which may be simultaneously or previously determined and/or contained in a database, to determine whether there is a match between them. Based on such information, the identity of the population of cells may be determined, or the identity of the population of cells may be distinguished from other populations of cells or cell types. Methods for determining a replication timing fingerprint for particular cell types are also provided.
摘要翻译：提供了基于使用高分辨率基因组阵列或测序程序的复制域时序概况来识别和/或区分同源细胞群的方法。这些方法可用于将细胞群体的复制定时概况与复制定时指纹，复制定时指纹和/或复制定时指纹的一个或多个信息段进行比较，其可以同时或先前确定和/或包含在数据库中，以确定它们之间是否存在匹配。基于这样的信息，可以确定细胞群体的身份，或者细胞群体的身份可以与其他细胞群体或细胞类型区分开。还提供了用于确定特定细胞类型的复制定时指纹的方法。

14. 发明授权

US06916613B2 Compositions, methods, kits and apparatus for determining the presence or absence of target molecules 失效
标题翻译：用于确定靶分子存在或不存在的组合物，方法，试剂盒和装置
公开(公告)号：US06916613B2
公开(公告)日：2005-07-12
申请号：US09844935
申请日：2001-04-27
申请人： Alexander Munishkin , Abraham Grossman
发明人： Alexander Munishkin , Abraham Grossman
IPC分类号： C12Q1/68 , C07H21/04 , C12N1/20 , C12N15/00 , C12N15/63
CPC分类号： C12Q1/6811 , C12Q2531/149
摘要： The present invention is directed to methods, compositions, kits and apparatus to identify and detect the presence or absence of target analytes. The embodiments of the present invention have utility in medical diagnosis and analysis of various chemical compounds in specimens and samples, as well as the design of test kits and apparatus for implementing such methods.
摘要翻译：本发明涉及用于鉴定和检测目标分析物的存在或不存在的方法，组合物，试剂盒和装置。本发明的实施例在标本和样品中的各种化学化合物的医学诊断和分析中具有实用性，以及用于实施这些方法的测试试剂盒和设备的设计。

15. 发明授权

US06699693B1 Process for DNA replication 失效
标题翻译： DNA复制过程
公开(公告)号：US06699693B1
公开(公告)日：2004-03-02
申请号：US09890829
申请日：2001-10-26
申请人： Kenneth Marians , Joing Liu
发明人： Kenneth Marians , Joing Liu
IPC分类号： C12P1934
CPC分类号： C12Q1/6853 , C12Q2531/149 , C12Q2525/301
摘要： A method is provided for replicating DNA, and in particular for replicating large segments of DNA. A primer is combined with a target DNA molecule to be replicated. The primer is designed to be at least partially homologous to a known site on the target DNA, and to create a D-loop when hybridized with that site. A replisome is then assembled at the D-loop, and this replisome creates a copy of the DNA, starting at the primer binding site. By utilizing two species of D-loop primers which bind to remote sites on the DNA flanking a region to be replicated, large sections of DNA can be replicated in a manner comparable to PCR. The replicated DNA can be analyzed to detect variations in the genetic sequence of the target, for linkage mapping and as a source of longer DNA molecules having a desired sequence.
摘要翻译：提供了用于复制DNA的方法，特别是用于复制大片段DNA的方法。将引物与待复制的靶DNA分子组合。引物设计为与目标DNA上的已知位点至少部分同源，并在与该位点杂交时产生D环。然后在D环上组装一个重复序列，并且该重复序列从引物结合位点开始创建DNA的拷贝。通过利用与待复制区域侧翼的DNA上的远端位点结合的两种D环引物，可以以与PCR相当的方式复制大部分DNA。可以分析复制的DNA以检测靶的遗传序列的变化，用于连锁作图和作为具有期望序列的较长DNA分子的来源。

16. 发明申请

US20020098485A1 Method for nucleic acid detection via Qbeta replicase 审中-公开
标题翻译：通过Qbeta复制酶检测核酸的方法
公开(公告)号：US20020098485A1
公开(公告)日：2002-07-25
申请号：US09781106
申请日：2001-02-08
发明人： Ann M. Morello , Qingping Jiang , John E. Monahan , Say-Jong Law
IPC分类号： C12Q001/68 , C12P019/34
CPC分类号： C12Q1/682 , C12Q1/6867 , C12Q2549/113 , C12Q2531/149 , C12Q2525/161
摘要： Disclosed is amplification of a target sequence using the activity of a replicase that quasi-autocatalytically replicates a specific replicase replicable or nullreplicatablenull sequence, while ensuring fidelity thereof by detecting the presence of the amplified target or antitarget sequence rather than the amplified replicase replicable sequence. A method according to the invention for assaying a target nucleic acid comprising hybridizing a set of one or more amplification probes with a nucleic acid sample is provided. A set of two chemiluminescent detection probes each for detecting portions of amplified target sequence along with a dual photomultiplier tube detection system for simultaneous detection of the two chemiluminescent detection probes are also provided for practicing the invention, for use with a target specific set of amplification probes. Kits for practicing the invention are also provided.
摘要翻译：公开了使用准自动复制特异性复制可复制或“可复制”序列的复制酶的活性扩增靶序列，同时通过检测扩增的靶或抗靶序列的存在而不是扩增的复制子可复制序列来确保其保真度。提供了根据本发明的用于测定靶核酸的方法，其包括将一组一个或多个扩增探针与核酸样品杂交。还提供了一组两个用于检测扩增的靶序列的部分以及用于同时检测两种化学发光检测探针的双光电倍增管检测系统的化学发光检测探针，用于实施本发明，用于靶特异性扩增探针组。还提供了用于实施本发明的试剂盒。

17. 发明授权

US06369207B1 Nucleic acid amplification with DNA-dependent RNA polymerase activity of RNA replicases 失效
公开(公告)号：US06369207B1
公开(公告)日：2002-04-09
申请号：US09396001
申请日：1999-09-14
申请人： Randall L. Dimond , Steven J. Ekenberg , James R. Hartnett , Geoffrey R. Hudson , Leopoldo G. Mendoza , Katharine M. Miller , John E. Monahan , Christopher L. Jones , Mark A. Maffitt , Richard A. Martinelli , Edward E. Pahuski , James W. Schumm
发明人： Randall L. Dimond , Steven J. Ekenberg , James R. Hartnett , Geoffrey R. Hudson , Leopoldo G. Mendoza , Katharine M. Miller , John E. Monahan , Christopher L. Jones , Mark A. Maffitt , Richard A. Martinelli , Edward E. Pahuski , James W. Schumm
IPC分类号： C07H1900
CPC分类号： C12Q1/6867 , C12Q1/6806 , C12Q1/6853 , C12Q2521/501 , C12Q2521/325 , C12Q2521/119 , C12Q2521/107 , C12Q2563/137 , C12Q2531/149 , C12Q2525/121
摘要： The present invention entails methods, and kits for carrying them out, based on the discovery that an RNA replicase, such as Q&bgr; replicase, has DNA-dependent RNA polymerase (“DDRP”) activity with nucleic acid segments, including DNA segments and DNA:RNA chimeric segments, which comprise a 2′-deoxyribonucleotide or an analog thereof and which have sequences of RNAs that are autocatalytically replicatable by the replicase. The discovery of this DDRP activity provides methods of the invention for nucleic acid amplification wherein a nucleic acid, with a DNA segment with the sequence of an RNA that is autocatalytically replicatable by an RNA replicase, is provided as a substrate for the replicase. The replicase catalyzes synthesis, from the DNA segment, of the RNA, which the replicase then autocatalytically replicates. The invention entails use of the amplification methods in detecting nucleic acid analytes, as in nucleic acid probe hybridization assays. Such assays of the invention include those wherein a nucleic acid analyte is hybridized with one or more nucleic acid probes, which include or are processed to generate a DNA segment which is amplifiable through production from the segment, catalyzed by the DDRP activity of an RNA replicase, of an autocatalytically replicatable RNA, which is autocatalytically replicated to provide an abundance of readily detectable reporter molecules. The invention permits replacement of an RNA, that is autocatalytically replicatable with an RNA replicase and employed as a reporter or label in prior art assays, such as nucleic acid probe hybridization assays or immunoassays, with a nucleic acid comprising a DNA segment with the same base sequence as the RNA. The invention also includes the methods of the invention with Mn+2, Co+2, or Zn+2 in the solutions in which the DDRP activity occurs.

18. 发明授权

US06297005B1 De novo priming activity of hepatitis C virus replicase 失效
标题翻译：丙型肝炎病毒复制酶的初始启动活性
公开(公告)号：US06297005B1
公开(公告)日：2001-10-02
申请号：US09305185
申请日：1999-05-04
申请人： Weidong Zhong , Zhi Hong , Annette Schettino Uss , Johnson Y. N. Lau
发明人： Weidong Zhong , Zhi Hong , Annette Schettino Uss , Johnson Y. N. Lau
IPC分类号： C12Q168
CPC分类号： C12Q1/6867 , C12N9/127 , C12Q1/707 , C12Q2525/186 , C12Q2531/149
摘要： The present invention relates to identification of a novel, de novo priming activity of hepatitis C virus replicase. This activity can be used to screen for anti-HCV replicase compounds, or to characterize the biological relevance of lead compounds that have already been identified.
摘要翻译：本发明涉及鉴定丙型肝炎病毒复制酶的新型引发活性。该活性可用于筛选抗HCV复制酶化合物，或表征已鉴定的铅化合物的生物相关性。

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