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1. 发明授权

US06524828B1 Mutant of RNA polymerases with increased stability 有权
标题翻译：增加稳定性的RNA聚合酶突变体
公开(公告)号：US06524828B1
公开(公告)日：2003-02-25
申请号：US09857475
申请日：2001-06-05
申请人： Hans Liao , Bob van Gemen , Akio Sugiyama
发明人： Hans Liao , Bob van Gemen , Akio Sugiyama
IPC分类号： C12P1934
CPC分类号： C12N9/1247 , C12Q1/6865
摘要： The present application relates to mutated RNA polymerases from bacteriophages that have increased stability, for example under high temperature conditions. One example of bacteriophage encoded RNA polymerase is the T7 RNA polymerase. T7 is a bacteriophage capable of infecting E. coli cells. Examples of other E. coli infecting T7-like bacteriophages are T3, øI, øII, W31, H, Y, A1, croC21, C22 and C23. An example of a Salmonella typhimurium infecting bacteriophage is SP6. The present invention is concerned with the RNA polymerases of T7-like bacteriophages that have been mutated. Due to these mutations the RNAP's have an increased stability. Preferred mutated RNA polymerases according to the invention are mutant RNA polymerases from T7 or SP3 bacteriophages. Due to the high homology between these enzymes, mutations in the T7 gene 1 sequence are likely to have the same effect in the corresponding gene sequence of the T3 bacteriophage. An especially preferred embodiment of the present invention is a T7 RNA polymerase with a serine to proline amino acid change in the protein at position 633 of the amino acid sequence. Since there is 80% homology between the T7 RNA polymerase and the T3 RNA polymerase the same effects of the 633 serine→proline mutation in the T7 gene may be expected for a 634 serine→proline amino acid mutation in the T3 RNA polymerase.
摘要翻译：本申请涉及来自具有增加的稳定性的噬菌体的突变的RNA聚合酶，例如在高温条件下。噬菌体编码的RNA聚合酶的一个实例是T7RNA聚合酶。 T7是能够感染大肠杆菌细胞的噬菌体。感染T7样噬菌体的其他大肠杆菌的实例是T3，øI，øII，W31，H，Y，A1，croC21，C22和C23。鼠伤寒沙门氏菌感染噬菌体的一个例子是SP6。本发明涉及已被突变的T7样噬菌体的RNA聚合酶。由于这些突变，RNAP具有增加的稳定性。根据本发明的优选的突变型RNA聚合酶是来自T7或SP3噬菌体的突变型RNA聚合酶。由于这些酶之间的高同源性，T7基因1序列中的突变可能在T3噬菌体的相应基因序列中具有相同的作用。本发明的特别优选的实施方案是具有丝氨酸以在氨基酸序列的633位的蛋白质中脯氨酸变化的T7RNA聚合酶。由于T7 RNA聚合酶和T3 RNA聚合酶之间具有80％的同源性，所以T3 RNA聚合酶中634丝氨酸 - >脯氨酸氨基酸突变可能预期T7基因中633丝氨酸 - >脯氨酸突变的相同作用。

2. 发明授权

US06967016B2 Method of determining therapeutic activity and/or possible side-effects of a medicament 有权
标题翻译：确定药物的治疗活性和/或可能的副作用的方法
公开(公告)号：US06967016B2
公开(公告)日：2005-11-22
申请号：US10006009
申请日：2001-12-04
申请人： Bob van Gemen , Eveline Catherina A. C. Timmermans , Anthonij de Ronde , Irene Johanna M. Dobbelaer
发明人： Bob van Gemen , Eveline Catherina A. C. Timmermans , Anthonij de Ronde , Irene Johanna M. Dobbelaer
IPC分类号： G01N33/50 , C12N15/09 , C12Q1/02 , C12Q1/68 , G01N33/15 , A61K49/00 , C12P19/34
CPC分类号： C12Q1/6888 , C12Q1/6809 , C12Q2600/136 , C12Q2600/158 , C12Q2600/16 , C12Q2545/101
摘要： The invention relates to the diagnosis of disease or the determination of functioning of cellular organisms, being of multi-cellular or unicellular nature, being visible by the naked eye or being a microorganism. The invention provides a method for determining functioning of a cellular organism comprising determining the relative ratio of a first endosymbiont cellular organelle nucleic acid and/or gene product thereof in a sample obtained from the organism in relation to the amount of a second nucleic acid and/or gene product thereof.
摘要翻译：本发明涉及诊断疾病或确定多细胞或单细胞性质的细胞生物的功能，由肉眼可视或是微生物。本发明提供了一种用于确定细胞生物功能的方法，包括确定从生物获得的样品中第一内共型细胞细胞器核酸和/或其基因产物相对于第二核酸的量和/ 或其基因产物。

3. 发明授权

US5834255A Quantification of nucleic acid 失效
标题翻译：核酸定量
公开(公告)号：US5834255A
公开(公告)日：1998-11-10
申请号：US385392
申请日：1995-02-07
申请人： Bob van Gemen , Tim Kievits , Peter F. Lens
发明人： Bob van Gemen , Tim Kievits , Peter F. Lens
IPC分类号： C07H21/04 , C12N15/09 , C12P19/34 , C12Q1/68 , C12Q1/70
CPC分类号： C12Q1/6851 , Y10S435/81
摘要： Disclosed is a method of quantifying a target nucleic acid in a test sample by adding to the test sample a known number of molecules of a corresponding nucleic acid comprising a well-defined mutant sequence. Said mutant sequence being discriminatory from the target nucleic acid. Subsequently a competitive amplification reaction of the nucleic acid is performed after which quantification of the amplified nucleic acid is performed by a differential detection.
摘要翻译：公开了一种通过向测试样品中加入已知数量的包含明确定义的突变序列的相应核酸分子的量化测试样品中的靶核酸的方法。所述突变序列与靶核酸区分开。随后进行核酸的竞争性扩增反应，之后通过差异检测进行扩增的核酸的定量。

4. 发明申请

US20060172326A1 HMG coenzyme a reductase inhibitors affect the mitochondrial DNA content of cells 审中-公开
标题翻译： HMG辅酶a还原酶抑制剂影响细胞的线粒体DNA含量
公开(公告)号：US20060172326A1
公开(公告)日：2006-08-03
申请号：US11321735
申请日：2005-12-29
申请人： Bob van Gemen , Marinus de Baar
发明人： Bob van Gemen , Marinus de Baar
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6883 , C12Q1/6851 , C12Q2600/16
摘要： Statins are widely used for the treatment of high cholesterol levels. In the present invention, the effect of statins on mitochondrial nucleic acid and the activation state of mitochondria is used in methods for determining whether a subject is at risk of developing side effects of statin treatment, methods for the treatment of a clinical symptom associated with reduced mitochondrial function. Further provided are kits and the like comprising a means for the detection of mitochondrial nucleic acid, or the activation of a mitochondrion for use in a method mentioned above.
摘要翻译：他汀类药物广泛用于治疗高胆固醇水平。在本发明中，他汀类药物对线粒体核酸和线粒体活化状态的作用用于确定受试者是否存在发生他汀类药物治疗副作用风险的方法，治疗与减少的临床症状相关的方法线粒体功能。进一步提供的试剂盒等包括用于检测线粒体核酸的方法或用于上述方法中的线粒体的活化。

5. 发明申请

US20050064478A1 Universal nucleic acid amplification system for nucleic acids in a sample 审中-公开
标题翻译：样品中核酸的通用核酸扩增系统
公开(公告)号：US20050064478A1
公开(公告)日：2005-03-24
申请号：US10915953
申请日：2004-08-10
申请人： Jing Zhang , John Dekker , Antoinette van der Kuyl , Jolanda Maas , Bob van Gemen
发明人： Jing Zhang , John Dekker , Antoinette van der Kuyl , Jolanda Maas , Bob van Gemen
IPC分类号： C07H19/00 , C07H21/00 , C07H21/04 , C12P19/34 , C12Q1/68 , C12Q1/6853 , C12Q1/70
CPC分类号： C12Q1/6853 , C12Q2531/113 , C12Q2525/179 , C12Q2525/155 , C12Q2525/143
摘要： Methods for amplifying nucleic acid in a sample including providing the sample with a set of primers to enable synthesis of at least one nucleic acid strand complementary to at least part of the nucleic acid, wherein the set of primers comprises between 3-8 random bases, preferably clustered near the 3′ end of each primer in said set of primers. The methods of the invention are useful, for example, for determining whether samples derived from humans, mammals, poultry, or fish comprise nucleic acid of a pathogen. The methods are further suited for typing the pathogen and typing particular variants of said pathogen. The methods are also suited for the elucidation of the gene expression profile or genetic profile of cells.
摘要翻译：用于在样品中扩增核酸的方法，包括向样品提供一组引物以使得能够合成与至少部分核酸互补的至少一个核酸链，其中所述引物组包含3-8个无规碱基，优选聚集在所述引物组中每个引物的3'末端附近。本发明的方法例如用于确定源自人，哺乳动物，家禽或鱼的样品是否包含病原体的核酸是有用的。所述方法还适用于分型病原体并分类所述病原体的特定变体。该方法也适用于阐明细胞的基因表达谱或遗传图谱。

6. 发明授权

US06750014B2 Reducing background in hybridization reactions 失效
标题翻译：减少杂交反应的背景
公开(公告)号：US06750014B2
公开(公告)日：2004-06-15
申请号：US09785881
申请日：2001-02-16
申请人： Marinus Petrus de Baar , Eveline Catherina A. C. Timmermans , Bob van Gemen
发明人： Marinus Petrus de Baar , Eveline Catherina A. C. Timmermans , Bob van Gemen
IPC分类号： C12Q168
CPC分类号： C12Q1/6827 , C12Q1/6832
摘要： The invention provides a method for reducing background in hybridization reactions of nucleic acids involving at least two homologous probes, wherein at least one of the probes is non-linear, or two homologous target sequences and a non-linear probe. Background is reduced by introducing an intended mismatch with a target sequence in at least one of the probes. The presence of the mismatch reduces the specificity of probes not entirely complementary to a target sequence to such an extent that the background signal is reduced. A set of mixed homologous probes, wherein at least one of the probes is non-linear, comprising such specific mismatch is also provided. The set can be used for the detection of variants of a family of nucleic acids, for instance a number of HIV variants. The invention also provides kits for carrying out the methods according to the invention.
摘要翻译：本发明提供了一种减少涉及至少两种同源探针的核酸的杂交反应背景的方法，其中至少一种探针是非线性的，或两个同源靶序列和非线性探针。通过在至少一个探针中引入目标序列的预期错配来减少背景。不匹配的存在降低了探针与靶序列不完全互补的特异性，使得背景信号降低的程度。还提供了一组混合同源探针，其中至少一个探针是非线性的，包括这种特异性错配。该组可用于检测核酸家族的变体，例如许多HIV变体。本发明还提供了用于实施根据本发明的方法的试剂盒。

7. 发明申请

US20050053963A1 Reducing background in hybridization reactions 审中-公开
标题翻译：减少杂交反应的背景
公开(公告)号：US20050053963A1
公开(公告)日：2005-03-10
申请号：US10751260
申请日：2004-01-02
申请人： Marinus de Baar , Eveline Timmermans , Bob van Gemen
发明人： Marinus de Baar , Eveline Timmermans , Bob van Gemen
IPC分类号： C12Q1/68 , C12Q1/6827 , C12Q1/6832
CPC分类号： C12Q1/6827 , C12Q1/6832
摘要： The invention provides a method for reducing background in hybridization reactions of nucleic acids involving at least two homologous probes, wherein at least one of the probes is nonlinear, or two homologous target sequences and a nonlinear probe. Background is reduced by introducing an intended mismatch with a target sequence in at least one of the probes. The presence of the mismatch reduces the specificity of probes not entirely complementary to a target sequence to such an extent that the background signal is reduced. A set of mixed homologous probes, wherein at least one of the probes is nonlinear, comprising such specific mismatch is also provided. The set can be used for the detection of variants of a family of nucleic acids, for instance, a number of HIV variants. The invention also provides kits for carrying out the methods according to the invention.
摘要翻译：本发明提供了减少涉及至少两个同源探针的核酸的杂交反应背景的方法，其中至少一个探针是非线性的，或两个同源靶序列和非线性探针。通过在至少一个探针中引入目标序列的预期错配来减少背景。不匹配的存在降低了探针与靶序列不完全互补的特异性，使得背景信号降低的程度。一组混合同源探针，其中至少一个探针是非线性的，还包括这种特异性失配。该组可用于检测核酸家族的变体，例如许多HIV变体。本发明还提供了用于实施根据本发明的方法的试剂盒。

8. 发明授权

US06808888B2 Universal nucleic acid amplification system for nucleic acids in a sample 失效
标题翻译：样品中核酸的通用核酸扩增系统
公开(公告)号：US06808888B2
公开(公告)日：2004-10-26
申请号：US10192786
申请日：2002-07-10
申请人： Jing Zhang , John Dekker , Antoinette C. van der Kuyl , Jolanda Maas , Bob van Gemen
发明人： Jing Zhang , John Dekker , Antoinette C. van der Kuyl , Jolanda Maas , Bob van Gemen
IPC分类号： C12Q168
CPC分类号： C12Q1/6853 , C12Q2531/113 , C12Q2525/179 , C12Q2525/155 , C12Q2525/143
摘要： Methods for amplifying nucleic acid in a sample comprising providing the sample with a set of primers to enable synthesis of at least one nucleic acid strand complementary to at least part of the nucleic acid, wherein the set of primers comprises between 3-8 random bases, preferably clustered near the 3′ end of each primer in said set of primers. The methods of the invention are useful, for example, for determining whether samples derived from humans, mammals, poultry, or fish comprise nucleic acid of a pathogen. The methods are further suited for typing the pathogen and typing particular variants of said pathogen. The methods are also suited for the elucidation of the gene expression profile or genetic profile of cells.
摘要翻译：用于在样品中扩增核酸的方法，包括向所述样品提供一组引物以使得能够合成至少一个与至少部分核酸互补的核酸链，其中所述引物组包含3-8个无规碱基，优选聚集在所述引物组中每个引物的3'末端附近。本发明的方法例如用于确定源自人，哺乳动物，家禽或鱼的样品是否包含病原体的核酸是有用的。所述方法还适用于分型病原体并分类所述病原体的特定变体。该方法也适用于阐明细胞的基因表达谱或遗传图谱。

9. 发明授权

US06110681A Primers and probes for the amplification, detection and typing of Mycoplasma pneumoniae 失效
标题翻译：用于扩增，检测和分型肺炎支原体的引物和探针
公开(公告)号：US06110681A
公开(公告)日：2000-08-29
申请号：US125324
申请日：1998-08-27
申请人： Caroline Louise Lucienne Ovyn , Bob van Gemen , Dianne Arnoldina Margaretha Wilhelmina van Strijp
发明人： Caroline Louise Lucienne Ovyn , Bob van Gemen , Dianne Arnoldina Margaretha Wilhelmina van Strijp
IPC分类号： C12N15/09 , C07H21/04 , C07K14/30 , C12P19/34 , C12Q1/68 , C12R1/35
CPC分类号： C12Q1/689 , C07K14/30
摘要： The present invention provides oligonucleotides that can be used as primers to amplify a region of the 16S rRNA of M. pneumoniae. Also provided are probes and kits for detection of amplified RNA and typing of M. pneumoniae strains. The primers, probes, methods and kits are useful for diagnosing M. pneumoniae.
摘要翻译： PCT No.PCT / EP97 / 00911 Sec。 371日期1998年8月27日第 102（e）日期1998年8月27日PCT 1997年2月25日PCT公布。出版物WO97 / 32036 PCT 日期1997年9月4日本发明提供可用作扩增肺炎支原体16S rRNA区域的引物的寡核苷酸。还提供了用于检测扩增的RNA和肺炎支原体菌株分型的探针和试剂盒。引物，探针，方法和试剂盒可用于诊断肺炎支原体。

10. 发明申请

US20080182260A1 Method for quantifying a ratio between at least two nucleic acid sequences 审中-公开
标题翻译：用于定量至少两个核酸序列之间的比率的方法
公开(公告)号：US20080182260A1
公开(公告)日：2008-07-31
申请号：US11985463
申请日：2007-11-15
申请人： Eveline C.A.C. Timmermans , Bob van Gemen
发明人： Eveline C.A.C. Timmermans , Bob van Gemen
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6851 , C12Q2527/113 , C12Q2545/101 , C12Q2545/113 , C12Q2561/101
摘要： The invention provides a method for quantifying an initial ratio of the amounts of at least two nucleic acids of interest in a sample by means of a multiplex nucleic acid amplification reaction, comprising amplifying the nucleic acids of interest in the amplification reaction, measuring the amount of at least two nucleic acids of interest at at least two different time points in the reaction, determining from at least two of the measurements the amplification rate of the at least two nucleic acids of interest, comparing the rates with a reference, and determining from the comparison the initial ratio of the amounts of the at least two nucleic acids of interest in the sample. Preferably, at least one variable factor in the nucleic acid amplification reaction is adjusted in order to allow detectable levels of all nucleic acids of interest to be reached before an amplification and/or detection limit of one or more of the nucleic acids of interest is reached. With the invention, small differences between ratios can be determined, as can direct measurement of the initial nucleic acid ratio. The invention is suitable for determining functioning of a cellular organism, the staging of a disease, and therapeutic activity and/or possible side effects of a compound.
摘要翻译：本发明提供了一种通过多重核酸扩增反应量化样品中至少两种感兴趣的核酸的量的初始比例的方法，包括扩增扩增反应中目的核酸，测量在所述反应中的至少两个不同时间点处至少两个感兴趣的核酸，从至少两个所述测量中确定所述至少两个感兴趣的核酸的扩增速率，将所述比率与参考值进行比较，并从比较样品中目的的至少两种核酸的量的初始比例。优选地，调节核酸扩增反应中的至少一个可变因子，以便在达到感兴趣的一种或多种感兴趣的核酸的扩增和/或检测限之前达到感兴趣的所有核酸的可检测水平。利用本发明，可以确定比率之间的小差异，直接测量初始核酸比例。本发明适合于确定细胞生物的功能，疾病的分期以及化合物的治疗活性和/或可能的副作用。

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