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公开(公告)号：US07148339B2
公开(公告)日：2006-12-12
申请号：US11088322
申请日：2005-03-23
摘要： The subject invention provides for a method of selectively cloning homoduplex nucleic acid molecules, in particular, by using a strain of host cells that contains a conditionally expressed and/or conditionally active mismatch-recognizing enzyme, e.g., a temperature sensitive variant of the gene encoding the endonuclease VII from phage T4. Using this host strain, the invention features a novel cloning method that selects for PCR products that are devoid of PCR-generated mutations.
摘要翻译： 本发明提供了选择性克隆同源双链核酸分子的方法，特别是通过使用含有条件表达和/或条件活性失配识别酶的宿主细胞株，例如编码基因的基因的温度敏感变体 来自噬菌体T4的核酸内切酶VII。 使用该宿主菌株，本发明的特征在于选择缺少PCR产生突变的PCR产物的新型克隆方法。
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公开(公告)号：US07118860B2
公开(公告)日：2006-10-10
申请号：US09728574
申请日：2000-11-30
申请人： Joseph A. Sorge , Anne M. Whalen
发明人： Joseph A. Sorge , Anne M. Whalen
CPC分类号： C12Q1/6823 , C12Q1/6816 , C12Q2521/301 , C12Q2531/113 , C12Q2521/307 , C12Q2525/161 , C12Q2525/301 , C12Q2561/109 , C12Q2565/519 , C12Q2561/101 , C12Q2521/107
摘要： The invention relates to a method of generating a signal indicative of the presence of a target nucleic acid in a sample, as well as a method of detecting or measuring a target nucleic acid in a sample. The invention comprises forming a cleavage structure by incubating a sample comprising a target nucleic acid with a probe having a secondary structure that changes upon binding of the probe to the target nucleic acid and further comprising a binding moiety. The invention also includes the steps of cleaving the cleavage structure with a nuclease to release a nucleic acid fragment to generate a signal, wherein generation of the signal is indicative of the presence of a target nucleic acid in a sample, and detecting and/or measuring the amount of the fragment captured by binding of a binding moiety to a capture element on a solid support.
摘要翻译： 本发明涉及产生指示样品中靶核酸存在的信号的方法，以及检测或测量样品中靶核酸的方法。 本发明包括通过将包含靶核酸的样品与具有二级结构的探针一起孵育形成切割结构，所述探针在探针与靶核酸结合后发生变化，并且还包含结合部分。 本发明还包括用核酸酶切割切割结构以释放核酸片段以产生信号的步骤，其中产生信号指示样品中靶核酸的存在，以及检测和/或测量 通过结合部分与固体支持物上的捕获元件结合捕获的片段的量。
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公开(公告)号：US07090978B2
公开(公告)日：2006-08-15
申请号：US10394485
申请日：2003-03-20
申请人： Manuel Perucho , Miguel Angel Peinado , Yurij Ionov , Sergei Malkhosyan , Michael McClelland , John Welsh
发明人： Manuel Perucho , Miguel Angel Peinado , Yurij Ionov , Sergei Malkhosyan , Michael McClelland , John Welsh
CPC分类号： C12Q1/6886 , C12Q2600/112 , C12Q2600/156
摘要： The detection of insertions and/or deletions in reiterated nucleotide sequences in tissues provides an identification of neoplastic changes that are associated with malignancy. The mutations are preferably detected by PCR based amplification of target sequences using selected primers, followed by standard analytic procedures. The detection of these mutations is useful as a diagnostic tool for cancer development and has direct application for cancer prognosis.
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84.发明授权公开(公告)号：US07078220B2
公开(公告)日：2006-07-18
申请号：US10800389
申请日：2004-03-12
IPC分类号： C12N1/20
摘要： The invention provided herein includes novel gram negative bacteria cells containing the Hte mutation. Other aspects of the invention include methods for rendering gram negative bacterial cells bearing the Hte region, such as E. coli cells competent for DNA transformation using any of a variety of competency inducing procedures. The competent cells of the subject invention may be frozen so as to provide for prolonged storage.
摘要翻译： 本文提供的本发明包括含有Hte突变的新型革兰氏阴性细菌细胞。 本发明的其它方面包括用于使用任何多种能力诱导程序提供具有Hte区域的革兰氏阴性细菌细胞的方法，例如能够进行DNA转化的大肠杆菌细胞。 本发明的感受态细胞可以被冷冻以提供长时间的储存。
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公开(公告)号：US20060099632A1
公开(公告)日：2006-05-11
申请号：US11300139
申请日：2005-12-14
申请人： Joseph Sorge
发明人： Joseph Sorge
CPC分类号： C12Q1/6844 , C12P19/34 , C12Q1/6813 , C12Q1/6869 , C12Q2525/125 , C12Q2521/301 , C12Q2521/331
摘要： The present invention provides a method of preparing a nucleic acid sample comprising template nucleic acid and synthetic nucleic acid for analysis wherein prior to analysis the nucleic acid sample is treated with a substance which selectively cleaves the template nucleic acid without substantially cleaving the synthetic nucleic acid. The invention further provides a method for improving the analysis of capillary-based DNA sequencing reactions, amplification reactions, and/or transcription reactions, wherein after the reaction, the nucleic acid sample comprising template nucleic acid and synthetic nucleic acid is treated with a substance which selectively cleaves the template without substantially cleaving the synthetic nucleic acid.
摘要翻译： 本发明提供了制备包含模板核酸和合成核酸用于分析的核酸样品的方法，其中在分析之前，用选择性切割模板核酸而不基本上切割合成核酸的物质处理核酸样品。 本发明进一步提供了改进基于毛细管的DNA测序反应，扩增反应和/或转录反应的分析方法，其中在反应之后，将包含模板核酸和合成核酸的核酸样品用 选择性地切割模板而基本上不裂合合成核酸。
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公开(公告)号：US20060014205A1
公开(公告)日：2006-01-19
申请号：US11230945
申请日：2005-09-19
申请人： Joseph Sorge , Rebecca Mullinax
发明人： Joseph Sorge , Rebecca Mullinax
CPC分类号： C12N9/1252 , C12Q1/686
摘要： The subject invention provides novel compositions containing a mixture of (a) an enzyme that possesses substantial 3′-5′ exonuclease activity (b) a DNA polymerase with less 3′-5′ exonuclease activity than the enzyme with substantial 3′-5′ exonuclease activity. Preferably, the DNA polymerase for inclusion in the compositions are DNA polymerases that substantially lack 3′-5′ exonuclease activity. A preferred embodiment of the invention is a composition comprising the Taq DNA polymerase (isolated from Thermus aquaticus) and the Pfu DNA polymerase (isolated from Pyrococcus furiosus). Another aspect of the invention is to provide methods for synthesizing polynucleotides, typically DNA, using compositions comprising an enzyme that possesses substantial 3′-5′ exonuclease activity and a DNA polymerase with less 3′-5′ exonuclease activity than the enzymes possessing substantial 3′-5′ exonuclease activity, preferably a DNA polymerase that substantially lacks 3′-5′ exonuclease activity. Another aspect of the invention involves the use the subject method of polynucleotide synthesis to carry out the synthesis step in a polymerase chain reaction experiment. Yet another aspect of the invention is to provide kits for the synthesis of polynucleotides, wherein the kits comprise an enzyme that possesses substantial 3′-5′ exonuclease activity and a DNA polymerase with less 3′-5′ exonuclease activity than the enzyme possessing substantial 3′-5′ exonuclease activity.
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公开(公告)号：US20050255512A1
公开(公告)日：2005-11-17
申请号：US11106237
申请日：2005-04-14
申请人： Joseph Sorge , Anne Whalen
发明人： Joseph Sorge , Anne Whalen
CPC分类号： C12Q1/6823 , C12Q1/6816 , C12Q2521/301 , C12Q2531/113 , C12Q2521/307 , C12Q2525/161 , C12Q2525/301 , C12Q2561/109 , C12Q2565/519 , C12Q2561/101 , C12Q2521/107
摘要： The invention relates to a method of generating a signal indicative of the presence of a target nucleic acid in a sample, where the method includes forming a cleavage structure by incubating a sample comprising a target nucleic acid with a probe having a secondary structure that changes upon binding of the probe to the target nucleic acid and further comprising a binding moiety. The invention also includes the steps of cleaving the cleavage structure with a nuclease to release a nucleic acid fragment to generate a signal, wherein generation of the signal is indicative of the presence of a target nucleic acid in a sample, and detecting and/or measuring the amount of the fragment captured by binding of a binding moiety to a capture element on a solid support. The invention also relates to a method of detecting or measuring a target nucleic acid in a sample, where the method includes forming a cleavage structure by incubating a sample containing a target nucleic acid with a probe having a secondary structure that changes upon binding of the probe to a target nucleic acid and comprising a binding moiety, and cleaving the cleavage structure with a nuclease to generate a cleaved nucleic acid fragment and detecting and/or measuring the amount of the fragment captured by binding of a binding moiety to a capture element on a solid support.
摘要翻译： 本发明涉及产生指示样品中目标核酸存在的信号的方法，其中该方法包括通过将包含靶核酸的样品与具有二级结构变化的探针一起孵育形成切割结构 探针与靶核酸的结合并且还包含结合部分。 本发明还包括用核酸酶切割切割结构以释放核酸片段以产生信号的步骤，其中产生信号指示样品中靶核酸的存在，以及检测和/或测量 通过结合部分与固体支持物上的捕获元件结合捕获的片段的量。 本发明还涉及一种检测或测量样品中的靶核酸的方法，其中该方法包括通过将含有靶核酸的样品与具有在探针结合后变化的二级结构的探针形成切割结构 并且包含结合部分，并用核酸酶切割切割结构以产生切割的核酸片段，并通过结合部分与捕获元件的结合捕获的片段的量，并检测和/或测量 坚实的支持。
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88.发明申请公开(公告)号：US20050184042A1
公开(公告)日：2005-08-25
申请号：US11103059
申请日：2005-04-11
申请人： Larry Brown , William Brumley , Kenneth Zajac
发明人： Larry Brown , William Brumley , Kenneth Zajac
IPC分类号： B01L3/00 , B01L7/00 , C12M1/00 , C12M1/38 , F27D11/00 , G05D23/20 , H02H3/05 , H05B3/06 , H05B3/20 , H05B3/34
CPC分类号： B01L3/50851 , B01L3/50853 , B01L7/52 , B01L7/54 , B01L2300/046 , B01L2300/0829 , B01L2300/1822 , B01L2300/1827 , B01L2300/1844
摘要： A flexible heating cover assembly for an apparatus for heating samples of biological material with substantial temperature uniformity includes a housing having a plurality of engageable enclosure components; a resistive heater having a plurality of heater element areas; a heater backing plate providing stability to the resistive heater; a force distribution system that distributes a force over the heater backing plate; and a support plate providing stiffness for the force distribution system, wherein the arrangement of the resistive heater, the heater backing plate, the force distribution system and the support plate provide substantial temperature uniformity among a plurality of sample tubes for receiving samples of biological material. The flexible heating cover assembly improves the uniformity, efficiency, quality, reliability and controllability of the thermal response during thermal cycling of the biological material.
摘要翻译： 一种用于加热具有显着温度均匀性的生物材料样品的装置的柔性加热盖组件包括具有多个可接合外壳部件的壳体; 电阻加热器，具有多个加热元件区域; 加热器背板，为电阻加热器提供稳定性; 力分配系统，其在加热器背板上分布力; 以及为力分配系统提供刚度的支撑板，其中电阻加热器，加热器背板，力分配系统和支撑板的布置在用于接收生物材料样品的多个样品管之间提供显着的温度均匀性。 柔性加热盖组件改善了生物材料热循环期间的热响应的均匀性，效率，质量，可靠性和可控性。
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公开(公告)号：US20050158711A1
公开(公告)日：2005-07-21
申请号：US10987388
申请日：2004-11-12
IPC分类号： A61K38/17 , C07H21/04 , C07K20060101 , C07K14/705 , C12N7/00 , C12N15/86 , C12Q1/68 , C12Q1/70
CPC分类号： C07H21/04
摘要： The invention provides for polynucleotides and vectors comprising at least two tag sequences. In particular, preferred vectors are viral vectors. The invention also provides for polynucleotides and vectors comprising a streptavidin binding peptide sequence and a calmodulin binding peptide sequence. The invention also provides for polynucleotides and vectors wherein a gene of interest is fused in frame to at least two tag sequences, for example, a streptavidin binding peptide sequence and a calmodulin binding peptide sequence. The invention also provides for methods of using the polynucleotides and vectors of the invention for detecting and/isolating protein complexes or identifying a binding partner for a protein of interest.
摘要翻译： 本发明提供了包含至少两个标签序列的多核苷酸和载体。 特别地，优选的载体是病毒载体。 本发明还提供了包含链霉亲和素结合肽序列和钙调蛋白结合肽序列的多核苷酸和载体。 本发明还提供多核苷酸和载体，其中目的基因与至少两个标签序列融合，例如链霉抗生物素蛋白结合肽序列和钙调蛋白结合肽序列。 本发明还提供了使用本发明的多核苷酸和载体用于检测和/或分离蛋白质复合物或鉴定感兴趣的蛋白质的结合配偶体的方法。
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90.发明申请Compositions and methods utilizing stable reporter cell lines for detection of pathway-specific signal transduction 审中-公开使用稳定的报道细胞系检测途径特异性信号转导的组合物和方法公开(公告)号：US20050106730A1
公开(公告)日：2005-05-19
申请号：US10818528
申请日：2004-04-05
申请人： Chao-Feng Zheng
发明人： Chao-Feng Zheng
CPC分类号： C12Q1/6897
摘要： The invention encompasses compositions and methods which utilize a cell line comprising a stably integrated recombinant nucleic acid construct comprising a reporter gene operably lined to a recognition sequence for a sequence-specific DNA-binding protein and a stably integrated recombinant nucleic acid construct comprising a sequence encoding a fusion protein, the fusion protein comprising a sequence-specific DNA binding domain, wherein the DNA binding domain specifically binds the recognition sequence, and a conditionally active transactivation domain, wherein activation of the conditionally active transactivation domain is dependent on protein phosphorylation and/or protein: protein interaction, and wherein binding of the fusion protein to the recognition sequence results in transactivation of the reporter gene expression when the transactivation domain fused to the DNA binding domain is activated.
摘要翻译： 本发明包括使用包含稳定整合的重组核酸构建体的细胞系的组合物和方法，所述重组核酸构建体包含可操作地排列于序列特异性DNA结合蛋白的识别序列的报道基因，以及稳定整合的重组核酸构建体，其包含编码 融合蛋白，所述融合蛋白包含序列特异性DNA结合结构域，其中所述DNA结合结构域特异性结合所述识别序列，以及条件活性反式激活结构域，其中所述条件活性反式激活结构域的活化取决于蛋白磷酸化和/或 蛋白质：蛋白质相互作用，并且当与DNA结合结构域融合的反式激活结构域被激活时，融合蛋白与识别序列的结合导致报告基因表达的反式激活。
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