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1. 发明授权

US08067205B2 Method of synthesizing cDNA 有权
标题翻译： cDNA合成方法
公开(公告)号：US08067205B2
公开(公告)日：2011-11-29
申请号：US10550788
申请日：2004-03-29
申请人： Seishi Kato , Tomoko Kimura , Kuniyo Ohtoko
发明人： Seishi Kato , Tomoko Kimura , Kuniyo Ohtoko
IPC分类号： C12P19/34
CPC分类号： C12N15/1096
摘要： A method for synthesizing cDNA possessing a consecutive sequence starting with a nucleotide adjacent to a cap structure of mRNA, which comprises (i) a process for annealing a double-stranded DNA primer and an RNA mixture containing mRNA possessing a cap structure, (ii) a process for preparing a conjugate of an mRNA/cDNA heteroduplex and a double-stranded DNA primer by synthesizing the first-strand cDNA primed with the double-stranded DNA primer using reverse transcriptase, and (iii) a process for circularizing the conjugate of the mRNA/cDNA heteroduplex and the double-stranded DNA primer by joining the 3′ and 5′ ends of the DNA strand containing cDNA using ligase. This method enables us to efficiently synthesize a full-length cDNA possessing a consecutive sequence starting with a transcription-start-site nucleotide from a small amount of RNA by small processes.
摘要翻译：一种合成cDNA的方法，该方法具有从与mRNA结构相邻的核苷酸开始的连续序列，其包括（i）退火双链DNA引物的方法和含有具有帽结构的mRNA的RNA混合物，（ii）通过使用逆转录酶合成用双链DNA引物引发的第一链cDNA来制备mRNA / cDNA异源双链和双链DNA引物的缀合物的方法，和（iii）使通过使用连接酶连接含有cDNA的DNA链的3'和5'末端，mRNA / cDNA异源双链和双链DNA引物。该方法使得我们能够通过小过程有效地合成具有从少量RNA转录起始位点核苷酸开始的连续序列的全长cDNA。

2. 发明申请

US20050053943A1 Human membrane antigen TM4 superfamily protein and DNA encoding this protein 审中-公开
标题翻译：人膜抗原TM4超家族蛋白和编码该蛋白的DNA
公开(公告)号：US20050053943A1
公开(公告)日：2005-03-10
申请号：US10608388
申请日：2003-06-30
申请人： Seishi Kato , Shingo Sekine , Tomoko Yamaguchi
发明人： Seishi Kato , Shingo Sekine , Tomoko Yamaguchi
IPC分类号： G01N33/53 , A61K38/00 , A61K48/00 , A61P35/00 , A61P43/00 , C07K14/47 , C07K14/705 , C12N15/09 , C12N15/12 , C12P21/02 , C12R1/19 , C12Q1/68 , C07H21/04 , C07K14/74
CPC分类号： C07K14/705 , A61K38/00
摘要： A human membrane antigen TM4 superfamily protein existing on the osteosarcoma cell surface and a cDNA encoding this protein is provided, said protein being useful as a pharmaceutical for treatment and diagnosis of cancers and also as an antigen for preparation of an antibody against said protein.
摘要翻译：提供存在于骨肉瘤细胞表面上的人膜抗原TM4超家族蛋白质和编码该蛋白质的cDNA，所述蛋白质可用作癌症治疗和诊断的药物，也可用作制备针对所述蛋白质的抗体的抗原。

3. 发明授权

US06280968B1 Human PEC-60-like protein and DNA encoding the same 失效
标题翻译：人类PEC-60样蛋白和编码相同的DNA
公开(公告)号：US06280968B1
公开(公告)日：2001-08-28
申请号：US09065019
申请日：1998-04-17
申请人： Seishi Kato , Tomoko Yamaguchi , Shingo Sekine , Kouju Kamata
发明人： Seishi Kato , Tomoko Yamaguchi , Shingo Sekine , Kouju Kamata
IPC分类号： C12P2106
CPC分类号： C07K14/575 , A61K38/00 , C07K2319/00
摘要： The present invention provides a human PEC-60-like protein, a gastrointestinal hormone excreted by a stomach tissue and a cDNA encoding this protein. The protein and the gene of the present invention can provide a protein containing the amino acid sequence represented by Sequence No. 1 and a DNA encoding said protein exemplified as a cDNA containing the base sequence represented by Sequence No. 1. as well as a human cDNA encoding a human PEC-60-like protein and said protein by the expression of this human cDNA recombinant.
摘要翻译：本发明提供人类PEC-60样蛋白质，由胃组织排出的胃肠激素和编码该蛋白质的cDNA。本发明的蛋白质和基因可以提供含有序列号1所示的氨基酸序列的蛋白质和编码所述蛋白质的DNA，该DNA作为含有序列号1表示的碱基序列的cDNA，以及人通过该人cDNA重组体的表达，编码人类PEC-60样蛋白质的cDNA和所述蛋白质。

4. 发明授权

US5597713A Process for producing cDNAs with complete length, process for producing intermediates thereof and process for producing vectors containing cDNAs with complete lengths 失效
标题翻译：用于生产具有完整长度的cDNA的方法，其制备中间体的方法和用于生产含有完整长度的cDNA的载体的方法
公开(公告)号：US5597713A
公开(公告)日：1997-01-28
申请号：US244188
申请日：1994-07-25
申请人： Seishi Kato , Shingo Sekine
发明人： Seishi Kato , Shingo Sekine
IPC分类号： C12N15/09 , C12N15/10 , C12N15/66 , C12N15/11 , C12N15/64 , C12P19/34
CPC分类号： C12N15/1096 , C12N15/66
摘要： A process for providing cDNAs containing full primary structure information of proteins by selectively synthesizing full-length cDNAs containing sequences starting from the capped region of mRNAs is disclosed. The invention provides a process for producing an intermediate for the synthesis of a full-length cDNA, which comprises the steps of treating mRNA extracted from cells to eliminate the phosphate group from the 5' end of an uncapped degraded mRNA; decapping from the 5' end of a capped intact mRNA; and ligating either a DNA oligonucleotide or a DNA-RNA chimeric oligonucleotide represented by the following general formula [I] to the 5' end phosphate group formed in the above step by the action of T4 RNA ligase, thereby selectively adding either the DNA oligonucleotide or the DNA-RNA chimeric oligonucleotide having an arbitrary sequence to the 5' end of the intact mRNA.5'-dN1-dN2- . . . -dNm-N1-N2- . . . -Nn-3' [I]The present invention also provides a process for synthesizing a full-length cDNA, which comprises linking a double-stranded DNA primer having a dT tail by annealing to a poly(A) tail of the 3' end of the intact mRNA having the DNA oligonucleotide or the DNA-RNA chimeric nucleotide added at the 5' end produced by the above-mentioned process, and then synthesizing the first strand cDNA complementary to the intact mRNA by a reverse transcriptase.
摘要翻译： PCT No.PCT / JP93 / 01359 Sec。 371日期：1994年7月25日 102（e）日期1994年7月25日PCT提交1993年9月22日PCT公布。第WO94 / 08001号公报日期1994年04月14日公开了通过选择性地合成含有从mRNA封端区开始的序列的全长cDNA提供含有蛋白质全部一级结构信息的cDNA的方法。本发明提供了一种制备用于合成全长cDNA的中间体的方法，其包括以下步骤：处理从细胞中提取的mRNA，以从未封端的降解的mRNA的5'末端消除磷酸基团; 从覆盖的完整mRNA的5'端开始切割; 并通过T4 RNA连接酶的作用将由以下通式[I]表示的DNA寡核苷酸或DNA-RNA嵌合寡核苷酸与上述步骤中形成的5'端磷酸基结合，从而选择性地添加DNA寡核苷酸或具有与完整mRNA的5'末端任意序列的DNA-RNA嵌合寡核苷酸。 5'-dN1-dN2-。。。 -dNm-N1-N2-。。。 -Nn-3'[I]本发明还提供了一种合成全长cDNA的方法，其包括将具有dT尾的双链DNA引物与3'端的poly（A）末端退火连接的具有通过上述方法制备的5'端加入的DNA寡核苷酸或DNA-RNA嵌合核苷酸的完整mRNA，然后通过逆转录酶合成与完整mRNA互补的第一链cDNA。

5. 发明申请

US20110207631A1 METHOD FOR PRODUCTION OF cDNA LIBRARY HAVING REDUCED CONTENT OF cDNA CLONE DERIVED FROM HIGHLY EXPRESSED GENE 审中-公开
标题翻译：用于生产具有由高表达基因衍生的cDNA克隆的还原内含子的cDNA文库的方法
公开(公告)号：US20110207631A1
公开(公告)日：2011-08-25
申请号：US13059927
申请日：2009-08-25
申请人： Kuniyo Ohtoko , Masahide Sugiyama , Seishi Kato
发明人： Kuniyo Ohtoko , Masahide Sugiyama , Seishi Kato
IPC分类号： C40B50/06 , C12N9/12 , C07H21/00
CPC分类号： C12N15/1096 , C12N15/1093 , C40B40/06 , C40B50/06 , C12Q2537/159
摘要： A method for efficiently constructing a cDNA library having a reduced content of cDNA clones derived from a highly expressed gene is provided. According to the method for constructing a cDNA library using a double-stranded DNA primer having oligo dT and mRNA as a template, the proportion of cDNA clones derived from a highly expressed gene in a cDNA library was decreased through coexistence with a probe having a property of binding to the mRNA of the highly expressed target gene so as to inhibit a cDNA extension reaction resulting from reverse transcriptase.
摘要翻译：提供了有效构建具有降低的来源于高表达基因的cDNA克隆含量降低的cDNA文库的方法。根据使用具有寡聚dT和mRNA作为模板的双链DNA引物构建cDNA文库的方法，通过与具有性质的探针共存，在cDNA文库中衍生自高表达基因的cDNA克隆的比例降低与高表达靶基因的mRNA结合，从而抑制由逆转录酶引起的cDNA延伸反应。

6. 发明申请

US20060246453A1 Method of synthesizing cdna 有权
标题翻译： cdna合成方法
公开(公告)号：US20060246453A1
公开(公告)日：2006-11-02
申请号：US10550788
申请日：2004-03-29
申请人： Seishi Kato , Tomoko Kimura , Kuniyo Ohtoko
发明人： Seishi Kato , Tomoko Kimura , Kuniyo Ohtoko
IPC分类号： C40B30/06 , C12Q1/68 , C07H21/02 , C07H21/04 , C12P19/34
CPC分类号： C12N15/1096
摘要： A method for synthesizing cDNA possessing a consecutive sequence starting with a nucleotide adjacent to a cap structure of mRNA, which comprises (i) a process for annealing a double-stranded DNA primer and an RNA mixture containing mRNA possessing a cap structure, (ii) a process for preparing a conjugate of an mRNA/cDNA heteroduplex and a double-stranded DNA primer by synthesizing the first-strand cDNA primed with the double-stranded DNA primer using reverse transcriptase, and (iii) a process for circularizing the conjugate of the mRNA/cDNA heteroduplex and the double-stranded DNA primer by joining the 3′ and 5′ ends of the DNA strand containing cDNA using ligase. This method enables us to efficiently synthesize a full-length cDNA possessing a consecutive sequence starting with a transcription-start-site nucleotide from a small amount of RNA by small processes.
摘要翻译：一种合成cDNA的方法，该方法具有从与mRNA结构相邻的核苷酸开始的连续序列，其包括（i）退火双链DNA引物的方法和含有具有帽结构的mRNA的RNA混合物，（ii）通过使用逆转录酶合成用双链DNA引物引发的第一链cDNA来制备mRNA / cDNA异源双链和双链DNA引物的缀合物的方法，和（iii）使通过使用连接酶连接含有cDNA的DNA链的3'和5'末端，mRNA / cDNA异源双链和双链DNA引物。该方法使得我们能够通过小过程有效地合成具有从少量RNA转录起始位点核苷酸开始的连续序列的全长cDNA。

7. 发明申请

US20050214734A1 Cell population provided with identification codes and method of screening cell population 审中-公开
标题翻译：细胞群体提供了筛选细胞群体的识别代码和方法
公开(公告)号：US20050214734A1
公开(公告)日：2005-09-29
申请号：US10509938
申请日：2003-04-01
申请人： Seishi Kato , Fumie Fujimori
发明人： Seishi Kato , Fumie Fujimori
IPC分类号： C12N5/10 , C12Q1/02 , G01N33/50 , C12N5/06 , C12Q1/00
CPC分类号： G01N33/5023 , G01N33/5008 , G01N33/502 , G01N33/5035
摘要： The invention of this application provides a cell population with identification codes, which is a population of cells that can be distinguished from one another based on a difference in luminescent signals emitted by luminescent substances, wherein the difference in the luminescent signals is caused by either or both (a) 2 or more different luminescent properties, and (b) 2 or more different luminescent sites. This cell population with identification codes can be conveniently prepared without a resort to a special apparatus and can be applied to either a solid phase system or a liquid phase system in mass screening, and moreover, can be used for a screening for various properties of cells such as the expression of a target protein.
摘要翻译：本申请的发明提供了具有识别码的细胞群，其是可以基于发光物质发射的发光信号的差异而彼此区分的细胞群，其中发光信号的差异是由任一种或（a）2种以上不同的发光性质，（b）2种以上不同的发光部位。具有识别码的细胞群可以方便地制备而无需使用特殊装置，并且可以在质谱筛选中应用于固相系统或液相系统，此外，可用于筛选细胞的各种性质例如靶蛋白的表达。

8. 发明授权

US6051424A Human cDNAs and proteins encoded thereby 失效
标题翻译：人类cDNA和由此编码的蛋白质
公开(公告)号：US6051424A
公开(公告)日：2000-04-18
申请号：US390207
申请日：1995-02-16
申请人： Seishi Kato , Suwan Oh , Shingo Sekine , Namsoon Kim , Takae Kato , Akiyo Iwahori
发明人： Seishi Kato , Suwan Oh , Shingo Sekine , Namsoon Kim , Takae Kato , Akiyo Iwahori
IPC分类号： C07K14/47 , C12N9/10 , C12N9/14 , C12N15/12 , C07H21/00 , C12N15/63
CPC分类号： C12N9/1007 , C07K14/47 , C12N9/14
摘要： Isolated cDNAs derived from mRNAs expressed in human cells are provided, as are DNAs and RNAs comprising their nucleotide sequences, and vectors for expressing the cDNAs. The cDNAs encode proteins which have functions similar to known proteins.
摘要翻译：提供了源自人细胞中表达的mRNA的分离的cDNA，以及包含其核苷酸序列的DNA和RNA以及用于表达cDNA的载体。 cDNA编码具有与已知蛋白质相似功能的蛋白质。

9. 发明授权

US5683898A Gene coding for eicosapentaenoic acid synthesizing enzymes and process for production of eicosapentaenoic acid 失效
标题翻译：二十碳五烯酸合成酶的基因编码和二十碳五烯酸的生产方法
公开(公告)号：US5683898A
公开(公告)日：1997-11-04
申请号：US375709
申请日：1995-01-20
申请人： Kazunaga Yazawa , Akiko Yamada , Seishi Kato , Kiyosi Kondo
发明人： Kazunaga Yazawa , Akiko Yamada , Seishi Kato , Kiyosi Kondo
IPC分类号： C12N1/21 , C12N9/00 , C12P7/64 , C07H19/00 , C12N1/20 , C12N15/00
CPC分类号： C12N9/93 , C12P7/6427 , C12P7/6472
摘要： There is provided an advantageous process for production of EPA by a gene recombinant technique wherein genes coding for biosynthesis enzymes for eicosapentaenoic acid (EPA) useful as pharmaceuticals, agrochemicals, foods, feeds or the like is obtained from microorganisms. EPA is produced by obtaining genes coding for eicosapentaenoic acid (EPA) biosynthesis enzymes, constructing a plasmid by joining the genes to a vector, transforming E. coli with the plasmid, and culturing the transformed E. coli.
摘要翻译：提供了通过基因重组技术生产EPA的有利方法，其中可从微生物获得编码二十碳五烯酸（EPA）的生物合成酶的基因，其用作药物，农业化学品，食品，饲料等。 EPA通过获得编码二十碳五烯酸（EPA）生物合成酶的基因产生，通过将基因连接到载体构建质粒，用质粒转化大肠杆菌，并培养转化的大肠杆菌。

10. 发明授权

US5624826A Cloning vector plasmid vector-primer derived therefrom and preparation method of the same 失效
标题翻译：来源于其的克隆载体质粒载体引物及其制备方法
公开(公告)号：US5624826A
公开(公告)日：1997-04-29
申请号：US466111
申请日：1995-06-06
申请人： Seishi Kato , Takashi Aoki , Yuri Umezawa
发明人： Seishi Kato , Takashi Aoki , Yuri Umezawa
IPC分类号： C12N15/00 , C07K14/525 , C12N15/09 , C12N15/10 , C12N15/70 , C12N15/85 , C12N15/64
CPC分类号： C12N15/1096 , C07K14/525 , C12N15/85 , C12N2800/108
摘要： Disclosed is a cloning vector plasmid suitable for preparing cDNA banks. The cloning vector plasmid of the present invention comprises, in the order mentioned from upstream to downstream region unless otherwise specified, a promoter PR1 acting in mammalian cells; an unique restriction enzyme site RE1 rarely existing in eukaryotic DNA and a promoter PR2 of RNA polymerase; an unique restriction enzyme site RE2 generating 3'-protruding dG tail; an unique arbitrary restriction enzyme site RE3; an unique arbitrary restriction enzyme site RE4 generating 3'-protruding terminus; an unique restriction enzyme site RE5 rarely existing in eukaryotic DNA and a promoter PR3 of RNA polymerase; and at least one kind of arbitrary restriction enzyme site RE6 generating 3'-protruding terminus existing at the arbitrary position before the restriction site RE1; and further, at arbitrary positions other than the above-described restriction enzyme sites and promoters, an origin OR1 for replication in mammalian cells, an origin OR2 for replication of a single-stranded phage, an origin OR3 for replication in E. coli, and a selective marker.
摘要翻译：公开了适用于制备cDNA库的克隆载体质粒。除非另有说明，本发明的克隆载体质粒以上游至下游区域的顺序包含在哺乳动物细胞中起作用的启动子PR1; 在真核DNA中很少存在的独特的限制酶位点RE1和RNA聚合酶的启动子PR2; 独特的限制酶位点RE2产生3'突出的dG尾巴; 独特的任意限制酶位点RE3; 独特的任意限制酶位点RE4产生3'突出的末端; 在真核DNA中很少存在一个独特的限制酶位点RE5和RNA聚合酶的启动子PR3; 以及存在于限制性位点RE1前的任意位置的产生3'突出末端的至少一种任意限制酶位点RE6; 此外，在除了上述限制酶位点和启动子之外的任意位置，用于在哺乳动物细胞中复制的起始OR1，用于复制单链噬菌体的起点OR2，用于在大肠杆菌中复制的起始OR3，以及选择性标记。

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