公开(公告)号：EP0870832A1

公开(公告)日：1998-10-14

申请号：EP96943336.6

申请日：1996-12-26

申请人： TAKARA SHUZO CO. LTD.

发明人： UEMORI, Takashi ,  ISHINO, Yoshizumi ,  KATO, Ikunoshin

IPC分类号： C12N15/54 ,  C12N9/12

CPC分类号： C12N9/1252

摘要： The present invention relates to a DNA polymerase possesses the properties of 1) exhibiting higher polymerase activity when assayed by using as a substrate a complex resulting from primer annealing to a single stranded template DNA, as compared to the case where an activated DNA is used as a substrate; 2) possessing a 3'→5' exonuclease activity; 3) being capable of amplifying a DNA fragment of about 20 kbp, in the case where polymerase chain reaction (PCR) is carried out using λ-DNA as a template. It also relates to a DNA polymerase-constituting protein; a DNA containing the base sequence encoding thereof; and a method for producing the DNA polymerase. The present invention provides a novel DNA polymerase possessing both a high primer extensibility and a 3'→5' exonuclease activity.

摘要翻译： 本发明涉及一种DNA聚合酶，具有以下特征：1）与使用活化DNA作为单链模板DNA的情况相比，使用由单链模板DNA引物退火得到的复合物作为底物进行测定时，具有较高的聚合酶活性 基材; 2）具有3'→5'核酸外切酶活性; 3）在使用λ-DNA作为模板进行聚合酶链反应（PCR）的情况下能够扩增约20kbp的DNA片段。 它还涉及构建DNA聚合酶的蛋白质; 含有编码其碱基序列的DNA; 以及DNA聚合酶的制造方法。 本发明提供具有高引物延伸性和3'-5'核酸外切酶活性的新型DNA聚合酶。

公开(公告)号：EP1167524A1

公开(公告)日：2002-01-02

申请号：EP00908056.5

申请日：2000-03-14

申请人： Takara Shuzo Co, Ltd.

发明人： MUKAI, Hiroyuki ,  YAMAMOTO, Junko ,  TAKEDA, Osamu ,  MIYAKE, Kazue ,  UEMORI, Takashi ,  SATO, Yoshimi ,  MORIYAMA, Mariko ,  SAWARAGI, Haruhisa ,  HAGIYA, Michio ,  ASADA, Kiyozo ,  KATO, Ikunoshin

IPC分类号： C12N15/10 ,  C12Q1/68

CPC分类号： C12Q1/6844 ,  C12Q2531/119 ,  C12Q2525/121 ,  C12Q2521/319

摘要： A convenient and effective method for amplifying a nucleic acid sequence characterized by effecting a DNA synthesis reaction in the presence of chimeric oligonucleotide primers; a method for supplying a large amount of DNA amplification fragments; an effective method for amplifying a nucleic acid sequence by combining the above method with another nucleic acid sequence amplification method; a method for detecting a nucleic acid sequence for detecting or quantitating a microorganism such as a virus, a bacterium, a fungus or a yeast; and a method for detecting a DNA amplification fragment obtained by the above method in situ.

摘要翻译： 一种用于扩增核酸序列的方法和方法，其特征在于在嵌合寡核苷酸引物存在下进行DNA合成反应; 提供大量DNA扩增片段的方法; 通过将上述方法与另一种核酸序列扩增方法结合来​​扩增核酸序列的有效方法; 用于检测用于检测或定量微生物如病毒，细菌，真菌或酵母的核酸序列的方法; 以及通过上述方法原位检测DNA扩增片段的方法。

公开(公告)号：EP1111044A1

公开(公告)日：2001-06-27

申请号：EP99940673.9

申请日：1999-09-06

申请人： Takara Shuzo Co, Ltd.

发明人： UEMORI, Takashi ,  SATO, Yoshimi ,  OKAWA, Mariko ,  FUJITA, Tomoko ,  MIYAKE, Kazue ,  TAKEDA, Osamu ,  SAGAWA, Hiroaki ,  HAGIYA, Michio ,  MUKAI, Hiroyuki ,  ASADA, Kiyozo ,  KATO, Ikunoshin

IPC分类号： C12N15/11 ,  C12Q1/68 ,  G01N33/53

CPC分类号： C12N15/1003 ,  C12Q1/686 ,  C12Q2527/125

摘要： A DNA synthesis method with a shortened time period required for DNA synthesis by polymerase chain reaction (PCR), characterized in that a DNA polymerase is used in an amount effective for providing more than 10 ng of amplified DNA fragments of about 2 kb per 50 µl of a reaction mixture, when PCR is carried out under the following conditions (A) and (B): (A) reaction mixture: 50 µl volume of a reaction mixture comprising DNA polymerase, 1 ng of genomic DNA from Escherichia coli, and 10 pmol each of primers Eco-1 and Eco-2 (nucleotide sequences of the primers Eco-1 and Eco-2 being shown in SEQ ID NOs: 10 and 11 of Sequence Listing, respectively); and having a composition suitable for the DNA polymerase; and (B) reaction conditions: 35 cycles of PCR, wherein one cycle consists of 99°C, 1 second - 66°C, 7 seconds; a kit for DNA synthesis usable for the DNA synthesis method; and an article of manufacture of a PCR agent. According to the present invention, the procedures in the genetic engineering studies and industries involved with PCR can be speeded up.

摘要翻译： 一种通过聚合酶链式反应（PCR）进行DNA合成所需的时间缩短的DNA合成方法，其特征在于使用DNA聚合酶，其用量可提供超过10ng的扩增DNA片段，每50ul约2kb （A）和（B）：（A）反应混合物：50μl体积的包含DNA聚合酶的反应混合物，1ng来自大肠杆菌的基因组DNA， 和引物Eco-1和Eco-2（引物Eco-1和Eco-2的核苷酸序列分别示于序列表的SEQ ID NO：10和11中）的10pmol; 并具有适合于DNA聚合酶的组合物; 和（B）反应条件：35个循环的PCR，其中一个循环由99℃，1秒-66℃，7秒; 用于DNA合成方法的DNA合成试剂盒; 和PCR试剂的制造。 根据本发明，可以加快基因工程研究中涉及PCR的工艺步骤。

公开(公告)号：EP0870839A1

公开(公告)日：1998-10-14

申请号：EP96937515.3

申请日：1996-11-07

申请人： TAKARA SHUZO CO. LTD.

发明人： ASADA, Kiyozo ,  UEMORI, Takashi ,  UENO, Takashi ,  KOYAMA, Nobuto ,  HASHINO, Kimikazu ,  KATO, Ikunoshiu

IPC分类号： C12N15/86 ,  C12N15/12 ,  C12N5/10

CPC分类号： C12N15/86 ,  A61K48/00 ,  C07K14/505 ,  C07K14/78 ,  C07K2319/00 ,  C12N9/1088 ,  C12N2740/13043 ,  C12N2740/13045

摘要： A method for increasing the efficiency of gene transfer into target cells with a retrovirus is disclosed. In the method, the target cells are infected with the retrovirus in the presence of either a mixture of an effective amount of a functional material having retrovirus binding domain and an effective amount of another functional material having target cell binding domain, or an effective amount of a functional material having these binding domains on the same molecule. The functional materials may be used without immobilization or with immobilized on beads. The method is useful, for example, gene therapy.

摘要翻译： 公开了一种提高逆转录病毒基因转移到靶细胞中的效率的方法。 在该方法中，目标细胞在有效量的具有逆转录病毒结合结构域的功能材料和有效量的具有靶细胞结合结构域的功能材料的混合物或有效量的 在同一分子上具有这些结合结构域的功能材料。 功能材料可以不固定地使用或固定在珠上。 该方法是有用的，例如基因治疗。

公开(公告)号：EP1072678A1

公开(公告)日：2001-01-31

申请号：EP99917081.4

申请日：1999-04-21

申请人： Takara Shuzo Co, Ltd.

发明人： ASADA, Kiyozo ,  UEMORI, Takashi ,  SATO, Yoshimi ,  FUJITA, Tomoko ,  MIYAKE, Kazue ,  TAKEDA, Osamu ,  MUKAI, Hiroyuki ,  KATO, Ikunoshin

IPC分类号： C12N15/10 ,  C12Q1/68

CPC分类号： C12P19/34 ,  C12N9/1252 ,  C12N9/96 ,  C12Q1/6806 ,  Y10S435/975 ,  C12Q2521/107

摘要： A DNA synthesis reaction-enhancer comprising at least one kind selected from the group consisting of acidic substances and cationic complexes; a DNA synthesis method in which during a DNA synthesis reaction a reaction is carried out in the presence of the above enhancer by using DNA polymerase; a DNA synthesis reaction composition comprising the above enhancer; a DNA synthesis reaction composition comprising two or more kinds of DNA polymerases each having 3 → 5 exonuclease activity; a DNA synthesis method in which during a DNA synthesis reaction two or more kinds of DNA polymerases each having 3 → 5 exonuclease activity are used; a kit for use in in vitro DNA synthesis, comprising two or more kinds of DNA polymerases each having 3 → 5 exonuclease activity; and a kit for use in in vitro DNA synthesis, wherein the kit comprises the DNA synthesis reaction-enhancer and DNA polymerase. According to the present invention, DNA synthesis can be carried out at an efficiency more excellent as compared to conventional DNA synthesis reaction.

摘要翻译： 一种DNA合成反应增强剂，其包含选自酸性物质和阳离子配合物中的至少一种; DNA合成方法，其中在DNA合成反应中，通过使用DNA聚合酶在上述增强子的存在下进行反应; 包含上述增强剂的DNA合成反应组合物; 一种DNA合成反应组合物，其包含两种或更多种各自具有3个→5个核酸外切酶活性的DNA聚合酶; 一种DNA合成方法，其中在DNA合成反应中使用两种或更多种具有3个→5个核酸外切酶活性的DNA聚合酶; 用于体外DNA合成的试剂盒，其包含两种或更多种各自具有3个→5个核酸外切酶活性的DNA聚合酶; 以及用于体外DNA合成的试剂盒，其中试剂盒包含DNA合成反应增强剂和DNA聚合酶。 根据本发明，与常规DNA合成反应相比，可以以更优异的效果进行DNA合成。

公开(公告)号：EP0997530A1

公开(公告)日：2000-05-03

申请号：EP98929687.6

申请日：1998-06-24

申请人： Takara Shuzo Co, Ltd.

发明人： UEMORI, Takashi ,  SATO, Yoshimi ,  FUJITA, Tomoko ,  MIYAKE, Kazue ,  MUKAI, Hiroyuki ,  ASADA, Kiyozo ,  KATO, Ikunoshin

IPC分类号： C12N15/54 ,  C12N9/12 ,  C12N15/31 ,  C07K14/195 ,  C12P21/02

CPC分类号： C12N9/1252

摘要： The present invention relates to a thermostable DNA polymerase-associated factor capable of enhancing DNA synthesizing-activity of a DNA polymerase; a thermostable DNA polymerase-associated factor possessing an activity of binding to a DNA polymerase and a method for producing the same; a gene encoding the DNA polymerase-associated factor; a method of DNA synthesis by using a DNA polymerase in the presence of the DNA polymerase-associated factor; and a kit comprising the DNA polymerase-associated factor. According to the present invention, there can be provided in vitro DNA synthesis and a DNA amplification system which are more excellent than conventional techniques by utilizing the DNA polymerase-associated factor of the present invention.

摘要翻译： 本发明涉及能够增强DNA聚合酶的DNA合成活性的耐热性DNA聚合酶相关因子; 具有与DNA聚合酶结合的活性的热稳定DNA聚合酶相关因子及其制备方法; 编码DNA聚合酶相关因子的基因; 在DNA聚合酶相关因子存在下使用DNA聚合酶进行DNA合成的方法; 和包含DNA聚合酶相关因子的试剂盒。 根据本发明，通过利用本发明的DNA聚合酶相关因子，可以提供比常规技术更优异的体外DNA合成和DNA扩增系统。