Enhancing the processivity of a family B-type DNA polymerase of Thermococcus onnurineus and application to long PCR
SCIE
SCOPUS
Cited 4 time in
WEB OF SCIENCE
Cited 4 time in
Scopus
-
Title
- Enhancing the processivity of a family B-type DNA polymerase of Thermococcus onnurineus and application to long PCR
-
Author(s)
- Kim, Yun Jae; Lee, Hyun Sook; Kwon, Suk-Tae; Lee, Jung-Hyun; Kang, Sung Gyun
- KIOST Author(s)
- Kim, Yun Jae(김윤재); Lee, Hyun Sook(이현숙); Lee, Jung Hyun(이정현); Kang, Sung Gyun(강성균)
-
Alternative Author(s)
- 김윤재; 이현숙; 이정현; 강성균
-
Publication Year
- 2014-05
-
Abstract
- Mechanisms that allow replicative DNA polymerases to attain high processivity are often specific to a given polymerase and cannot be generalised to others. Amplification efficiency is lower in family B-type DNA polymerases than in family A-type (Taq) polymerases because of their strong 3'aEuro"5' exonuclease-activity. Here, we have red the exonuclease domain of the Thermococcus onnurineus NA1 (TNA1) DNA polymerase, especially Asn210 to Asp215 residues in Exo II motif (NXXXFD), to improve the processivity. N213D mutant protein had higher processivity and extension rate than the wild-type TNA1 DNA polymerase, retaining a lower mutation frequency than recombinant Taq DNA polymerase. Consequently, the N213D mutant could amplify target DNA up to 13.5 kb in length from human genomic DNA and 16.2 kb in length from human mitochondrial DNA while wild-type TNA1 amplified target DNA of 2.7 kb in length from human genomic DNA.
-
ISSN
- 0141-5492
-
URI
- https://sciwatch.kiost.ac.kr/handle/2020.kiost/2828
-
DOI
- 10.1007/s10529-013-1441-x
-
Bibliographic Citation
- BIOTECHNOLOGY LETTERS, v.36, no.5, pp.985 - 992, 2014
-
Publisher
- SPRINGER
-
Subject
- HYPERTHERMOPHILIC ARCHAEON; EXONUCLEASE ACTIVITY; HIGH-FIDELITY; AMPLIFICATION; CLONING; LITORALIS; DUTPASE; ENZYME; SITE; NA1
-
Keywords
- DNA polymerase; Error rate; 3 ' -5 ' Exonuclease domain; Processivity; Thermococcus DNA polymerase; TNA1 DNA polymerase
-
Type
- Article
-
Language
- English
-
Document Type
- Article
- Files in This Item:
-
There are no files associated with this item.
Items in ScienceWatch@KIOST are protected by copyright, with all rights reserved, unless otherwise indicated.