公开(公告)号：US20050069887A1

公开(公告)日：2005-03-31

申请号：US10495581

申请日：2002-11-14

申请人： Masao Kitabayashi ,  Toshihiro Kuroita ,  Yoshikazu Ishida ,  Hideyuki Komatsubara ,  Yoshiaki Nishiya ,  Masanori Oka ,  Yoshihisa Kawamura ,  Tadayuki Imanaka

发明人： Masao Kitabayashi ,  Toshihiro Kuroita ,  Yoshikazu Ishida ,  Hideyuki Komatsubara ,  Yoshiaki Nishiya ,  Masanori Oka ,  Yoshihisa Kawamura ,  Tadayuki Imanaka

IPC分类号： C07K14/195 ,  C12N9/12 ,  C12P19/34 ,  C12Q1/68

CPC分类号： C07K14/195 ,  C12N9/1252 ,  C12Q1/6846 ,  C12Q2527/125 ,  C12Q2521/101

摘要： A composition for enhancing synthesis of DNA comprising an anion-supplying substance that is effective in promoting DNA synthesis in enzymatic DNA synthesis reactions, in particular a salt of a dicarboxylic acid. Further enhanced DNA synthesis promoting effects can be achieved by using, together with the anion-supplying substance, at least one compound selected from the group consisting of dimethylsulfoxide and compounds represented by the following formula (1): R2—CH2—NR1xHy  (1) wherein R1 is an alkyl group having 1 to 3 carbon atoms, R2 is a substituent selected from the group consisting of the following (a) and (b): (a) oxygen and (b) radicals represented by the formula wherein R4 is methyl, hydrogen or forms a pyrrolidine ring when combined with R1, R5 is —CO2H or —SO3H, and n is an integer from 0 to 2, x is an integer from 1 to 3 and y is an integer from 0 to 2, provided that x plus y equals 3. The present invention further provides a thermostable DNA polymerase-related factor derived from Thermococcus species, which has an activity to promote the DNA synthesis activity of DNA polymerase or bind to DNA polymerase.

摘要翻译： 一种用于增强DNA合成的组合物，其包含在促进DNA合成反应中有效促进DNA合成的阴离子供给物质，特别是二羧酸的盐。 通过与阴离子供给物质一起使用至少一种选自二甲基亚砜和下式（1）表示的化合物的化合物，可以实现进一步增强的DNA合成促进效果：R 2 -CH 2 -NR <1> xHy（1）其中R 1是具有1至3个碳原子的烷基，R 2是选自以下（a）和（b）的取代基：（a）氧 和（b）由下式表示的基团其中R 4是甲基，氢或当与R 1一起形成吡咯烷环时，R 5是-CO 2 H或-SO 3 H，n是0至 2，x是1至3的整数，y是0至2的整数，条件是x加y等于3.本发明还提供了源自Thermococcus物种的热稳定DNA聚合酶相关因子，其具有活性 促进DNA聚合酶的DNA合成活性或与DNA聚合酶结合。

公开(公告)号：US6008025A

公开(公告)日：1999-12-28

申请号：US902632

申请日：1997-07-29

申请人： Hideyuki Komatsubara ,  Masao Kitabayashi ,  Hideki Kamimura ,  Bunsei Kawakami ,  Yoshihisa Kawamura ,  Masahiro Takagi ,  Tadayuki Imanaka

发明人： Hideyuki Komatsubara ,  Masao Kitabayashi ,  Hideki Kamimura ,  Bunsei Kawakami ,  Yoshihisa Kawamura ,  Masahiro Takagi ,  Tadayuki Imanaka

IPC分类号： C12N9/12 ,  C12N15/54 ,  C12Q1/68 ,  C12P19/34

CPC分类号： C12Q1/686 ,  C12N9/1252 ,  C12Q1/6848

摘要： A modified thermostable DNA polymerase having 5% or less of the 3'-5' exonuclease activity of the enzyme before modification and a DNA polymerase composition for amplifying nucleic acid, which comprises the modified thermostable DNA polymerase having 0 to 5% of the 3'-5' exonuclease activity of the enzyme before modification and an unmodified thermostable DNA polymerase having 3'-5' exonuclease activity or a modified thermostable DNA polymerase having 100 to 6% of the 3'-5' exonuclease activity of a thermostable DNA polymerase before modification; a method for amplifying nucleic acid by use of said modified thermostable polymerase or said DNA polymerase composition; and a reagent therefor.

摘要翻译： 一种修饰前的酶的3'5'核酸外切酶活性的5％以下的扩增型热稳定型DNA聚合酶和扩增核酸的DNA聚合酶组合物，其含有3' -5'核酸外切酶活性和具有3'-5'核酸外切酶活性的未修饰的热稳定DNA聚合酶或经热稳定DNA聚合酶的3'-5'核酸外切酶活性的修饰的热稳定DNA聚合酶的100-6％ 修改; 通过使用所述修饰的热稳定聚合酶或所述DNA聚合酶组合物来扩增核酸的方法; 和其试剂。

公开(公告)号：US07422882B2

公开(公告)日：2008-09-09

申请号：US09852922

申请日：2001-05-10

申请人： Toshihiro Kuroita ,  Masao Kitabayashi ,  Yoshikazu Ishida ,  Hideyuki Komatsubara ,  Yoshiaki Nishiya ,  Bunsei Kawakami ,  Yoshihisa Kawamura ,  Tadayuki Imanaka

发明人： Toshihiro Kuroita ,  Masao Kitabayashi ,  Yoshikazu Ishida ,  Hideyuki Komatsubara ,  Yoshiaki Nishiya ,  Bunsei Kawakami ,  Yoshihisa Kawamura ,  Tadayuki Imanaka

IPC分类号： C12N9/12 ,  C12P19/34

CPC分类号： C12N9/1252

摘要： An object of the present invention is to provide a thermostable DNA polymerase with enhanced amplification efficiency and/or improved fidelity in polymerase chain reaction (PCR), and provide a process for production thereof. More specifically, the present invention provides thermostable DNA polymerase wherein in the DX1EX2X3X4H sequence (D: aspartic acid, E: glutamic acid, H: histidine, X1, X2, X3 and X4: any amino acid) consisting of DX1E sequence within the EXO I region and a four amino acid length peptide adjacent to said glutamic acid(E) of thermostable DNA polymerase having 3′-5′ exonuclease activity, histidine(H) has been replaced by another amino acid.

摘要翻译： 本发明的目的是提供具有增强的扩增效率和/或聚合酶链反应（PCR）中的改进的保真度的热稳定性DNA聚合酶，并提供其生产方法。 更具体地说，本发明提供了热稳定的DNA聚合酶，其中在X 1，X 2，X 3，X 4， 序列（D：天冬氨酸，E：谷氨酸，H：组氨酸，X 1，X 2，X 3和X 3， 由SEQ ID NO：1区域内的DX1E序列组成的4个氨基酸长度肽与邻近所述谷氨酸（E）的4个氨基酸长度的肽组成，所述热稳定性DNA聚合酶具有3' -5'核酸外切酶活性，组氨酸（H）已被另一种氨基酸取代。

公开(公告)号：US07384739B2

公开(公告)日：2008-06-10

申请号：US10495581

申请日：2002-11-14

申请人： Masao Kitabayashi ,  Toshihiro Kuroita ,  Yoshikazu Ishida ,  Hideyuki Komatsubara ,  Yoshiaki Nishiya ,  Masanori Oka ,  Yoshihisa Kawamura ,  Tadayuki Imanaka

发明人： Masao Kitabayashi ,  Toshihiro Kuroita ,  Yoshikazu Ishida ,  Hideyuki Komatsubara ,  Yoshiaki Nishiya ,  Masanori Oka ,  Yoshihisa Kawamura ,  Tadayuki Imanaka

IPC分类号： C12Q1/68 ,  C12P19/34 ,  C12N9/00 ,  C08B3/00

CPC分类号： C07K14/195 ,  C12N9/1252 ,  C12Q1/6846 ,  C12Q2527/125 ,  C12Q2521/101

摘要： The invention provides methods, kits, and compositions for enhancing synthesis of DNA involving a carboxylate ion-supplying substance that is effective in promoting DNA synthesis in enzymatic DNA synthesis reactions. The invention further provides a thermostable DNA polymerase-related factor derived from Thermococcus species, which has an activity to promote the DNA synthesis activity of DNA polymerase or which binds to DNA polymerase.

摘要翻译： 本发明提供用于增强DNA合成的方法，试剂盒和组合物，所述方法，试剂盒和组合物涉及在酶促DNA合成反应中有效促进DNA合成的羧酸根离子供应物质。 本发明还提供了源自Thermococcus种的热稳定DNA聚合酶相关因子，其具有促进DNA聚合酶的DNA合成活性或与DNA聚合酶结合的活性。