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1. 发明授权

US07659060B2 Method for identifying nucleotide polymorphism 有权
标题翻译：识别核苷酸多态性的方法
公开(公告)号：US07659060B2
公开(公告)日：2010-02-09
申请号：US10653586
申请日：2003-09-02
申请人： Yutaka Takarada , Yoshihiro Soya , Yoshihisa Kawamura
发明人： Yutaka Takarada , Yoshihiro Soya , Yoshihisa Kawamura
IPC分类号： C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6827 , C12Q2565/101 , C12Q2563/173
摘要： A method for identifying a nucleotide polymorphism, the method comprising hybridizing a labeled oligonucleotide for a wild type or a mutant type to a nucleic acid containing a specific nucleotide polymorphic site in a sample; allowing a nucleic acid-specific label to act thereon; detecting an interaction between the label of the oligonucleotide and the nucleic acid-specific label; and identifying a nucleotide polymorphism, wherein the identification utilizes a difference due to the nucleotide polymorphism in conditions under which the hybridized oligonucleotide is separated into a single strand.
摘要翻译：一种用于鉴定核苷酸多态性的方法，所述方法包括将野生型或突变型的标记寡核苷酸与含有样品中特定核苷酸多态性位点的核酸杂交; 允许核酸特异性标记作用于其上; 检测寡核苷酸的标记与核酸特异性标记之间的相互作用; 并鉴定核苷酸多态性，其中鉴定在杂交寡核苷酸被分离成单链的条件下利用由于核苷酸多态性引起的差异。

2. 发明授权

US5981183A Method for amplifying and detecting of target nucleic acid sequence using thermostable enzyme 失效
标题翻译：使用热稳定酶扩增和检测靶核酸序列的方法
公开(公告)号：US5981183A
公开(公告)日：1999-11-09
申请号：US821782
申请日：1997-03-20
申请人： Yutaka Takarada , Hiroaki Inoue , Shuji Shibata , Yoshihisa Kawamura
发明人： Yutaka Takarada , Hiroaki Inoue , Shuji Shibata , Yoshihisa Kawamura
IPC分类号： C12Q1/68 , C07H21/04 , C12P19/34
CPC分类号： C12Q1/6865
摘要： In a method for amplifying the specific nucleic acid sequence, a highly specific amplification which has low possibility of non-specific hybridization can be carried out. Highly stable reagents, the activities of which do not decrease in case of supply and storage, are also provided. Thermostable enzymes are used as RNA dependent DNA polymerase, DNA dependent DNA polymerase, DNA dependent RNA polymerase and ribonuclease H, which are required for the amplification system based on replicated RNA. Especially, it is preferable that a thermostable enzyme derived from Thermus thermophilus which has RNA dependent DNA polymerase activity, DNA dependent DNA polymerase activity and ribonuclease H activity, and thermostable DNA dependent RNA polymerase are used together. By this method, inactivation of enzymes are prevented by using thermostable enzymes, and the amplification can be carried out without sequential addition of enzymes.
摘要翻译：在扩增特异性核酸序列的方法中，可以进行非特异性杂交的可能性低的高度特异性扩增。还提供了稳定的试剂，其供应和储存的活性不降低。热稳定酶用作基于复制RNA的扩增系统所需的RNA依赖性DNA聚合酶，DNA依赖性DNA聚合酶，DNA依赖性RNA聚合酶和核糖核酸酶H。特别优选共同使用具有RNA依赖性DNA聚合酶活性，DNA依赖性DNA聚合酶活性和核糖核酸酶H活性的嗜热栖热菌的热稳定性酶和耐热性DNA依赖型RNA聚合酶。通过这种方法，通过使用热稳定酶来防止酶的失活，并且可以不依次加入酶进行扩增。

3. 发明授权

US06303306B1 Method for amplifying and detecting of target nucleic acid sequence using thermostable enzyme 有权
标题翻译：使用热稳定酶扩增和检测靶核酸序列的方法
公开(公告)号：US06303306B1
公开(公告)日：2001-10-16
申请号：US09292435
申请日：1999-04-15
申请人： Yutaka Takarada , Hiroaki Inoue , Shuji Shibata , Yoshihisa Kawamura
发明人： Yutaka Takarada , Hiroaki Inoue , Shuji Shibata , Yoshihisa Kawamura
IPC分类号： C12Q168
CPC分类号： C12Q1/6865 , C12Q2527/101
摘要： In a method for amplifying the specific nucleic acid sequence, a highly specific amplification which has low possibility of non-specific hybridization can be carried out. Highly stable reagents, the activities of which do not decrease in case of supply and storage, are also provided. Thermostable enzymes are used as RNA dependent DNA polymerase, DNA dependent DNA polymerase, DNA dependent RNA polymerase and ribonuclease H, which are required for the amplification system based on replicated RNA. Especially, it is preferable that a thermostable enzyme derived from Thermus thermophilus which has RNA dependent DNA polymerase activity, DNA dependent DNA polymerase activity and ribonuclease H activity, and thermostable DNA dependent RNA polymerase are used together. By this method, inactivation of enzymes are prevented by using thermostable enzymes, and the amplification can be carried out without sequential addition of enzymes.
摘要翻译：在扩增特异性核酸序列的方法中，可以进行非特异性杂交的可能性低的高度特异性扩增。还提供了稳定的试剂，其供应和储存的活性不降低。热稳定酶用作基于复制RNA的扩增系统所需的RNA依赖性DNA聚合酶，DNA依赖性DNA聚合酶，DNA依赖性RNA聚合酶和核糖核酸酶H。特别优选共同使用具有RNA依赖性DNA聚合酶活性，DNA依赖性DNA聚合酶活性和核糖核酸酶H活性的嗜热栖热菌的热稳定性酶和耐热性DNA依赖型RNA聚合酶。通过这种方法，通过使用热稳定酶来防止酶的失活，并且可以不依次加入酶进行扩增。

4. 发明授权

US07601504B2 Protein achieving improved blocking efficiency 失效
标题翻译：蛋白质提高了阻塞效率
公开(公告)号：US07601504B2
公开(公告)日：2009-10-13
申请号：US10562776
申请日：2004-07-02
申请人： Toshihiro Kuroita , Atsushi Sogabe , Yutaka Takarada , Naoki Tanaka
发明人： Toshihiro Kuroita , Atsushi Sogabe , Yutaka Takarada , Naoki Tanaka
IPC分类号： G01N33/53
CPC分类号： C07K14/245 , G01N33/54393
摘要： It is intended to provide a method of conveniently screening a novel protein or a novel partial sequence protein having a blocking ability based on amino acid sequence data; and a protein achieving an improved blocking efficiency that can be expressed on a large scale in Escherichia coli. A method of screening a novel protein or a novel partial sequence protein having a blocking ability based on amino acid sequence data; a protein characterized by achieving an improved blocking efficiency owing to an amino acid sequence modification; and a method of utilizing the protein.
摘要翻译：旨在提供一种方便地筛选具有基于氨基酸序列数据的阻断能力的新型蛋白质或新型部分序列蛋白质的方法; 和能够在大肠杆菌中大规模表达的提高的阻断效率的蛋白质。一种基于氨基酸序列数据筛选具有阻断能力的新型蛋白质或新型部分序列蛋白质的方法; 一种蛋白质，其特征在于由于氨基酸序列修饰而获得改善的阻断效率; 以及利用该蛋白质的方法。

5. 发明授权

US06582922B1 Method of extracting nucleic acids using particulate carrier 失效
标题翻译：使用颗粒载体提取核酸的方法
公开(公告)号：US06582922B1
公开(公告)日：2003-06-24
申请号：US10067208
申请日：2002-02-07
申请人： Katsuya Daimon , Shigeru Komai , Yutaka Takarada
发明人： Katsuya Daimon , Shigeru Komai , Yutaka Takarada
IPC分类号： C12Q168
CPC分类号： C12N15/1006 , C12N15/1013 , C12Q1/6806 , C12Q2565/137
摘要： The present invention provides a method of extracting and isolating nucleic acids from a material containing nucleic acids using a nucleic acid-binding particulate carrier. More specifically, the present invention provides a nucleic acid extraction method using a particulate carrier having a particle diameter of 0.5 to 15.0 &mgr;m, a pore diameter of 50 to 500 nm and a pore volume of 200 to 5000 mm3/g. According to the method of the invention, nucleic acids can be efficiently extracted from a biological material, in particular a material containing a large amount of contaminants, such as a clinical sample.
摘要翻译：本发明提供了使用核酸结合颗粒载体从含有核酸的材料中提取和分离核酸的方法。更具体地说，本发明提供使用粒径为0.5〜15.0μm，孔径为50〜500nm，孔体积为200〜5000mm 3 / g的粒状载体的核酸提取方法。根据本发明的方法，可以从生物材料，特别是含有大量污染物的材料（例如临床样品）中有效地提取核酸。

6. 发明申请

US20060183173A1 Protein achieving improved blocking efficiency 失效
公开(公告)号：US20060183173A1
公开(公告)日：2006-08-17
申请号：US10562776
申请日：2004-07-02
申请人： Toshihiro Kuroita , Atsushi Sogabe , Yutaka Takarada , Naoki Tanaka
发明人： Toshihiro Kuroita , Atsushi Sogabe , Yutaka Takarada , Naoki Tanaka
IPC分类号： G01N33/537 , G01N33/53 , G01N33/543
CPC分类号： C07K14/245 , G01N33/54393
摘要： It is intended to provide a method of conveniently screening a novel protein or a novel partial sequence protein having a blocking ability based on amino acid sequence data; and a protein achieving an improved blocking efficiency that can be expressed on a large scale in Escherichia coli. A method of screening a novel protein or a novel partial sequence protein having a blocking ability based on amino acid sequence data; a protein characterized by achieving an improved blocking efficiency owing to an amino acid sequence modification; and a method of utilizing the protein.

7. 发明申请

US20070292853A1 Dna Array and Method for Detecting Single Nucleotide Polymorphism 审中-公开
标题翻译： Dna阵列和检测单核苷酸多态性的方法
公开(公告)号：US20070292853A1
公开(公告)日：2007-12-20
申请号：US10593349
申请日：2005-03-18
申请人： Mitsuo Kawase , Yasuko Yoshida , Kazunari Yamada , Yutaka Takarada , Kouzo Hashimoto
发明人： Mitsuo Kawase , Yasuko Yoshida , Kazunari Yamada , Yutaka Takarada , Kouzo Hashimoto
IPC分类号： C07H21/04 , C12Q1/68
CPC分类号： C12Q1/6827 , C12Q2565/501 , C12Q2537/143 , C12Q2525/204
摘要： A DNA array for detecting a single nucleotide polymorphism of a gene, which comprises, on a solid support, a first probe spots group consisting of one or more probe spots each containing one or more probes hybridizable with a polynucleotide of the gene, in which the probes are exactly complementary to the first polymorphism pattern of the gene, and a second probe spots group consisting of one or more probe spots each containing one or more probes hybridizable with the polynucleotide of the gene, in which the probes are exactly complementary to the second polymorphism pattern of the gene, wherein probe lengths in the probe spots is different from each other. This DNA array enables more exact SNP detection.
摘要翻译：一种用于检测基因的单核苷酸多态性的DNA阵列，其在固体支持物上包含由一个或多个探针点组成的第一探针斑点，每个探针点含有一个或多个与该基因的多核苷酸杂交的探针，其中探针与基因的第一多态性模式完全互补，第二探针点组由一个或多个探针点组成，每个探针点含有一个或多个可与基因的多核苷酸杂交的探针，其中探针与第二个基因的多态性模式，其中探针点中的探针长度彼此不同。该DNA阵列能够进行更准确的SNP检测。

8. 发明授权

US06281008B1 Nucleic acid extraction apparatus 失效
标题翻译：核酸提取装置
公开(公告)号：US06281008B1
公开(公告)日：2001-08-28
申请号：US09241242
申请日：1999-02-01
申请人： Shigeru Komai , Katsuya Daimon , Yutaka Takarada
发明人： Shigeru Komai , Katsuya Daimon , Yutaka Takarada
IPC分类号： C12M300
CPC分类号： B01L3/502 , B01L3/5082 , B01L2200/026 , B01L2300/042 , B01L2300/046 , B01L2300/049 , B01L2400/0487 , B01L2400/0622 , B01L2400/0644 , C12N15/1013 , G01N1/405 , G01N35/0098 , G01N2035/0436 , Y10S435/82
摘要： This invention provides an apparatus for extracting nucleic acids from nucleic acid-containing samples. The nucleic acid extraction apparatus of the invention comprises (1) a group of extraction vessels each comprising a reactor tube in which a sample, a reagent solution, and a magnetic carrier are admixed and reacted, a drain cup for pooling an unwanted component solution, and a nucleic acid recovery tube all as secured to a support, (2) a distribution means for introducing a solution into each of the extraction vessels, (3) a stirring means for mixing the solution and magnetic carrier in the reactor tube, (4) a holding means for holding the magnetic carrier stationary within the vessel, (5) a discharging means for discharging the solution from the reactor tube while the magnetic carrier is held stationary, (6) a heating means for heating the solution and magnetic carrier in the reactor tube, and (7) a transfer means for serially transferring the vessels to the given positions.
摘要翻译：本发明提供了一种从含核酸样品中提取核酸的装置。本发明的核酸提取装置包括（1）一组提取容器，每个提取容器均包含反应器管，其中将样品，试剂溶液和磁性载体混合并反应，用于汇集不需要的组分溶液的排水杯，（2）用于将溶液引入每个提取容器的分配装置，（3）用于将溶液和磁性载体混合在反应器管中的搅拌装置，（4））用于将磁性载体固定在容器内的保持装置，（5）一个放电装置，用于在磁性载体保持静止时将溶液从反应器管排出;（6）加热装置，用于将溶液和磁性载体加热反应器管，和（7）用于将容器串联转移到给定位置的转移装置。

9. 发明授权

US5525462A Nucleic acid sequence amplification method, detection method, and reagent kit therefor 失效
标题翻译：核酸序列扩增法，检测方法及试剂盒
公开(公告)号：US5525462A
公开(公告)日：1996-06-11
申请号：US875758
申请日：1992-04-28
申请人： Yutaka Takarada , Toshiya Aono , Shuji Shibata
发明人： Yutaka Takarada , Toshiya Aono , Shuji Shibata
IPC分类号： C12Q1/68 , C07H21/04 , C12P19/34
CPC分类号： C12Q1/6853
摘要： An oligonucleotide is provided, which has at least a base sequence A, which is 10-30 nucleotides in length and homologous to a part of a specific nucleic acid sequence to be amplified, and a base sequence B, which is 10-30 nucleotides in length and complementary to a sequence 3' to the part of the specific nucleic acid sequence to be amplified. Sequences A and B are separated by a spacer region of 0-20 nucleotides. The oligonucleotide is used in an amplification method, wherein the oligonucleotide is mixed with a sample containing a specific nucleic acid sequence to be amplified and allowed to anneal to the specific nucleic acid sequence in the sample. The annealed oligonucleotide acts as a primer in an elongation reaction, whereas any nonannealed oligonucleotide is decomposed, except for at least part of the base sequence A. The elongation product is then denatured to a single strand for use as a template to which the remaining oligonucleotide anneals to form a primer for subsequent elongation. Also provided is a detection method, which allows for detection of a specific nucleic acid sequence in a sample, as well as reagent kits for use in the amplification and detection methods.
摘要翻译：提供了寡核苷酸，其至少具有碱基序列A，其长度为10-30个核苷酸并且与待扩增的特异性核酸序列的一部分同源;以及碱基序列B，其为10-30个核苷酸长度并与要扩增的特定核酸序列部分的序列3'互补。序列A和B被0-20个核苷酸的间隔区隔开。寡核苷酸用于扩增方法，其中将寡核苷酸与含有待扩增的特异性核酸序列的样品混合并使其与样品中的特定核酸序列退火。退火的寡核苷酸在伸长反应中用作引物，而除了至少部分碱基序列A之外，任何非退火的寡核苷酸都被分解。然后将延伸产物变性为单链，用作剩余的寡核苷酸退火以形成随后伸长的底漆。还提供了一种检测方法，其允许检测样品中的特定核酸序列，以及用于扩增和检测方法的试剂盒。

10. 发明授权

US06261773B1 Reagent for nucleic acid amplification and process for nucleic acid amplification 失效
标题翻译：用于核酸扩增的试剂和用于核酸扩增的方法
公开(公告)号：US06261773B1
公开(公告)日：2001-07-17
申请号：US09268710
申请日：1999-03-16
申请人： Masaya Segawa , Motohiro Kondo , Yutaka Takarada
发明人： Masaya Segawa , Motohiro Kondo , Yutaka Takarada
IPC分类号： C12Q168
CPC分类号： C12Q1/6848 , C12Q2527/125
摘要： The present invention provide a process for sequence-specific nucleic acid amplification capable of improving the detection sensitivity and increasing the signal. In particular, the present invention provides a reagent for nucleic acid amplification containing at least one member selected from the group consisting of EDTA, NTA, UDA, CyDTA, DTPA, GEDTA, TTHA and their salts, specifically a reagent for nucleic acid amplification comprising, in addition to the at least one compound, a forward primer having a DNA sequence homologous to a sequence of a target RNA; a reverse primer having a DNA sequence complementary to a sequence of the target RNA and having a promoter for RNA polymerase attached to its 5′ end; ribonucleotides; deoxyribonucleotides; a reverse transcriptase or RNA-directed DNA polymerase; a RNase H; a DNA polymerase or a reverse transcriptase having DNA-directed DNA polymerase activity; and a RNA polymerase. The present invention also provides a process for nucleic acid amplification characterized by carrying out a nucleic acid amplification reaction in the presence of at least one member selected from the group consisting of EDTA, NTA, UDA, CyDTA, DTPA, GEDTA, TTHA and their salts.
摘要翻译：本发明提供能够提高检测灵敏度和增加信号的序列特异性核酸扩增方法。特别地，本发明提供了一种用于核酸扩增的试剂，其包含选自EDTA，NTA，UDA，CyDTA，DTPA，GEDTA，TTHA及其盐中的至少一种，特别是用于核酸扩增的试剂，除了至少一种化合物之外，具有与靶RNA的序列同源的DNA序列的正向引物; 具有与靶RNA序列互补的DNA序列并具有连接于其5'末端的RNA聚合酶的启动子的反向引物; 核糖核苷酸脱氧核糖核苷酸; 逆转录酶或RNA指导的DNA聚合酶; 核糖核酸酶H; 具有DNA定向DNA聚合酶活性的DNA聚合酶或逆转录酶; 和RNA聚合酶。本发明还提供了一种核酸扩增方法，其特征在于在选自EDTA，NTA，UDA，CyDTA，DTPA，GEDTA，TTHA及其盐中的至少一种的存在下进行核酸扩增反应。

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