公开(公告)号：US20100221787A1

公开(公告)日：2010-09-02

申请号：US12739496

申请日：2008-10-24

申请人： Yoshihide Hayashizaki ,  Masayoshi Itoh ,  Alexander Lezhava ,  Yoshimi Benno ,  Yasumasa Mitani ,  Hajime Kanamori

发明人： Yoshihide Hayashizaki ,  Masayoshi Itoh ,  Alexander Lezhava ,  Yoshimi Benno ,  Yasumasa Mitani ,  Hajime Kanamori

IPC分类号： C12P19/34 ,  C12N9/12

CPC分类号： C12Q1/6846 ,  C12N9/1252 ,  C12P19/34 ,  C12Q2525/155 ,  C12Q2521/107 ,  C12Q2527/101 ,  C12Q2531/107

摘要： A DNA polymerase suitable for specific isothermal amplification methods and an isothermal amplification method using the DNA polymerase are provided. In the presence of a DNA polymerase including a protein described in the following item (a) or (b), an amplification reaction of a target nucleic acid sequence in a nucleic acid sample is carried out isothermally using a first primer shown in the following (X). By using the DNA polymerase, it becomes possible to carry out the amplification reaction using the primer within a shorter time than ever before. (a) a protein having an amino acid sequence represented by SEQ ID NO. 23 (b) a protein having an amino acid sequence represented by SEQ ID NO. 25 (X) a primer that contains, in a 3′ end portion, a sequence (Ac′) that hybridizes to a sequence (A) of a 3′ end portion of the target nucleic acid sequence and also contains, on a 5′ side of the sequence (Ac′), a sequence (B′) that hybridizes to a complementary sequence (Bc) to a sequence (B) present on a 5′ side with respect to the sequence (A) in the target nucleic acid sequence

摘要翻译： 提供了适用于特定等温扩增方法的DNA聚合酶和使用DNA聚合酶的等温扩增方法。 在包含下述（a）或（b）中所述的蛋白质的DNA聚合酶的存在下，使用下述第一引物等温进行靶核酸序列的扩增反应 X）。 通过使用DNA聚合酶，可以在比以往更短的时间内使用引物进行扩增反应。 （a）具有SEQ ID NO：1所示的氨基酸序列的蛋白质。 23（b）具有SEQ ID NO：1所示的氨基酸序列的蛋白质。 25（X）引物，其在3'末端含有与目标核酸序列的3'端部分的序列（A）杂交的序列（Ac'），并且在5' 序列（Ac'）侧，与靶序列（A）相对于存在于5'侧的序列（B）的互补序列（Bc）杂交的序列（B'）

公开(公告)号：US20100047862A1

公开(公告)日：2010-02-25

申请号：US12083695

申请日：2006-10-20

申请人： Yoshihide Hayashizaki ,  Masayoshi Itoh ,  Yoshimi Benno ,  Alexander Lezhava

发明人： Yoshihide Hayashizaki ,  Masayoshi Itoh ,  Yoshimi Benno ,  Alexander Lezhava

IPC分类号： C12P21/00 ,  C12N9/12 ,  C07H21/00 ,  C12N15/63 ,  C12N1/00 ,  C12P19/34

CPC分类号： C12N9/1276 ,  C12N9/1252

摘要： This invention provides a novel DNA polymerase obtained from Bacillus smithii JCM9076, which has novel features in terms of, for example, optimal reaction conditions (e.g., optimal temperature) and enzyme activity. More particularly, a novel DNA polymerase is a pol I type DNA polymerase, which is any of proteins (a) to (f) below and has DNA polymerase activity: (a) a protein comprising the amino acid sequence as shown in SEQ ID NO: 7; (b) a protein consisting of an amino acid sequence derived from the amino acid sequence as shown in SEQ ID NO: 7 by deletion, substitution, or addition of one or several amino acid residues; (c) a protein consisting of the amino acid sequence as shown in SEQ ID NO: 9; and (d) a protein consisting of an amino acid sequence derived from the amino acid sequence as shown in SEQ ID NO: 9 by deletion, substitution, or addition of one or several amino acid residues.

摘要翻译： 本发明提供了一种从芽孢杆菌JCM9076获得的新型DNA聚合酶，其在例如最佳反应条件（例如最佳温度）和酶活性方面具有新特征。 更具体地，新型DNA聚合酶是pol I型DNA聚合酶，其是以下蛋白质（a）至（f）中的任一种，并且具有DNA聚合酶活性：（a）包含SEQ ID NO所示的氨基酸序列的蛋白质 ：7; （b）通过缺失，取代或添加一个或几个氨基酸残基的由SEQ ID NO：7所示的氨基酸序列衍生的氨基酸序列组成的蛋白质; （c）由SEQ ID NO：9所示的氨基酸序列组成的蛋白质; 和（d）通过缺失，取代或添加一个或几个氨基酸残基，由SEQ ID NO：9所示的氨基酸序列衍生的氨基酸序列组成的蛋白质。