公开(公告)号：KR100673836B1

公开(公告)日：2007-01-24

申请号：KR1020050107202

申请日：2005-11-09

Applicant: 성균관대학교산학협력단

Inventor： 권석태 ,  서무석

IPC: C12N15/31 ,  C12N15/52 ,  C12N15/09

Abstract: A novel Staphylothermus marinus DNA ligase is provided to be thermostable and be used for various reactions for catalyzing phosphodiester bond formation in a nucleic acid. The DNA ligase includes an amino acid sequence described as SEQ ID : NO. 2. The method for preparing the DNA ligase comprises the steps of: (a) preparing a vector expressing the DNA ligase; (b) transforming the vector into a host cell; (c) culturing the transformant; and (d) recovering protein from the transformant.

Abstract translation: 提供了一种新颖的葡萄球菌DNA连接酶，其是热稳定的，并用于催化核酸中磷酸二酯键形成的各种反应。 DNA连接酶包括SEQ ID NO：1所示的氨基酸序列。 制备DNA连接酶的方法包括以下步骤：（a）制备表达DNA连接酶的载体; （b）将载体转化为宿主细胞; （c）培养转化体; 和（d）从转化体中回收蛋白质。

公开(公告)号：KR100774102B1

公开(公告)日：2007-11-06

申请号：KR1020060125956

申请日：2006-12-12

Applicant: 성균관대학교산학협력단

Inventor： 권석태 ,  서무석 ,  이정현

IPC: C12N15/57 ,  C12N15/09

Abstract: A thermostable enzyme DNA ligase derived from Sulfophobococcus zilligii is provided to improve resistance against strong acid or alkali condition, and against low or high temperature condition, so that the enzyme is useful in various reactions for promoting formation of a phosphodiester linkage in nucleic acids. A thermostable DNA ligase derived from Sulfophobococcus zilligii, Szi DNA ligase, has the amino acid sequence of SEQ ID NO:2, employs ATP and ADP as cofactors and Mg^2+ and Mn^2+ as metal cofactors, and shows optimum activity at pH 6-7, more than 60% activity at 70-80 deg.C, and more than 50% activity at 100 deg.C for 2.2 hours. A gene encoding the thermostable DNA ligase derived from Sulfophobococcus zilligii has the nucleotide sequence of SEQ ID NO:1. A recombinant vector pESZL(KACC 95099P) contains the thermostable DNA ligase gene. The thermostable DNA ligase is produced by transforming a host cell with the recombinant vector pESZL(KACC 95099P), culturing the transformed cell, and recovering the enzyme from the transformed cell.

Abstract translation: 提供源自磺化磷酸链球菌的热稳定酶DNA连接酶以提高对强酸或碱性条件的抗性，并且在低或高温条件下，使酶可用于促进在核酸中形成磷酸二酯键的各种反应。 源自Sulfophobococcus zilligii，Szi DNA连接酶的热稳定DNA连接酶具有SEQ ID NO：2的氨基酸序列，使用ATP和ADP作为辅因子，Mg 2+和Mn 2+作为金属辅因子，并且在 pH6-7，70-80℃下超过60％活性，在100℃下活性超过50％，持续2.2小时。 编码源自磺化磷酸链球菌的热稳定DNA连接酶的基因具有SEQ ID NO：1的核苷酸序列。 重组载体pESZL（KACC 95099P）含有热稳定DNA连接酶基因。 通过用重组载体pESZL（KACC 95099P）转化宿主细胞，培养转化的细胞并从转化的细胞中回收酶来产生热稳定DNA连接酶。