公开(公告)号：KR1020100107873A

公开(公告)日：2010-10-06

申请号：KR1020090026203

申请日：2009-03-27

Applicant: 성균관대학교산학협력단

Inventor： 권석태 ,  배희진 ,  김기쁨

IPC: C12N9/12 ,  C12N15/63

Abstract: PURPOSE: A thermal resistant DNA polymerase which is derived from Thermococcus celer with DNA polymerization activity and proofreading activity is provided to widely use in various nucleic acid polymerization. CONSTITUTION: A thermal resistant Tce DNA polymerase isolated from Thermococcus celer contains an amino acid of seuqnce number 10. The Tce DNA polymerase has an optimal activity in 20-35 mM of (NH_4)_2SO_4 and 4-16 mM of Mg^2+ at pH 6.5 and 75°C. A nucleic acid molecule encoding the Tce DNA polymerase contains a nucleic acid sequence of sequence number 9. A pTCE vector contains a nucleic acid molecule encoding the Tce DNA polymerase. A method for preparing the Tce DNA polymerase comprises: a step of preparing a vector expressing the Tce DNA polymerase; a step of transforming the vector to the microorganism; a step of culturing the transformant; and a step of collecting protein from the transformant.

Abstract translation: 目的：提供具有DNA聚合活性和校正活性的源自Thermococcus celer的耐热DNA聚合酶，广泛用于各种核酸聚合。 构成：从Thermococcus celer分离的耐热Tce DNA聚合酶含有第10号氨基酸.Tce DNA聚合酶在20-35mM（NH_4）_2SO_4和4-16mM Mg2 +中具有最佳活性 pH 6.5和75℃。 编码Tce DNA聚合酶的核酸分子含有序列号9的核酸序列.PTCE载体含有编码Tce DNA聚合酶的核酸分子。 制备Tce DNA聚合酶的方法包括：制备表达Tce DNA聚合酶的载体的步骤; 将载体转化到微生物的步骤; 培养转化体的一个步骤; 以及从转化体中收集蛋白质的步骤。

公开(公告)号：KR101241769B1

公开(公告)日：2013-03-25

申请号：KR1020110012680

申请日：2011-02-14

Applicant: 성균관대학교산학협력단

Inventor： 권석태 ,  김기쁨

IPC: C12N9/12 ,  C12N15/54 ,  C12N1/21 ,  C12Q1/68

Abstract: 본 발명은 DNA 중합 활성(DNA polymerization activity) 및 교정 활성 (proofreading activity)을 모두 가지는 써모코커스 씰러리크레센스 유래의 DNA 중합효소(
Thermococcus

celericrescens
DNA polymerase,
Tcel DNA polymerase, 이하 '
Tcel DNA 중합효소'라고 함) 및 상기
Tcel DNA 중합효소에서 점 돌연변이에 의하여 752번째 아미노산인 알라닌이 라이신으로, 213번째 아미노산인 아스파라진이 아스팔틱산으로 이중 치환된 돌연변이 써모코커스 씰러리크레센스 A752K/N213D DNA 중합효소 (
Thermococcus

celericrescens
A752K/N213D DNA polymerase,
Tcel A752K/N213D DNA polymerase, 이하 '
Tcel A752K/N213D DNA 중합효소'라고 함)에 관한 것이다.

公开(公告)号：KR1020120092820A

公开(公告)日：2012-08-22

申请号：KR1020110012680

申请日：2011-02-14

Applicant: 성균관대학교산학협력단

Inventor： 권석태 ,  김기쁨

IPC: C12N9/12 ,  C12N15/54 ,  C12N1/21 ,  C12Q1/68

CPC classification number: C12N9/1252 ,  C12Q1/686 ,  C12Q2521/101 ,  C12R1/01

Abstract: PURPOSE: A Thermococcus celericrescens-derived DNA polymerase having DNA polymerization activity and proofreading activity is provided to enable various nucleic acid polymerization reactions and to improved PCR efficiency and thermal resistance. CONSTITUTION: A Tcel DNA polymerase isolated from Thermococcus celericrescens has an amino acid sequence of sequence number 14. A nucleic acid molecule encoding the polymerase has a base of sequence number 13. A recombinant vector contains the nucleic acid molecule. TcelA752K/N213D DNA polymerase has an optimal activity at pH 8, 70 Deg. C. 100 mM of KCl, and 2 mM or Mg^2+. A recombinant vector, pTCELM vector, contains the nucleic acid molecule encoding the TcelA752K/N213D DNA polymerase. A microorganism transformed by the vector is E.coli Rosetta(DE3)pLysS / pTCELM (deposit number KACC91614P).

Abstract translation: 目的：提供具有DNA聚合活性和校正活性的Thermococcus celericrescens衍生的DNA聚合酶，以实现各种核酸聚合反应并提高PCR效率和耐热性。 构成：从Thermococcus celericrescens分离的Tcel DNA聚合酶具有序列号14的氨基酸序列。编码聚合酶的核酸分子具有序列号13的碱基。重组载体含有核酸分子。 TcelA752K / N213D DNA聚合酶在pH8,70度下具有最佳活性。 100mM KCl和2mM或Mg 2+。 重组载体pTCELM载体含有编码TcelA752K / N213D DNA聚合酶的核酸分子。 由载体转化的微生物是大肠杆菌Rosetta（DE3）pLysS / pTCELM（保藏号KACC91614P）。