Cloning, expression, and partial characterization of a family B-type DNA polymerase from the hyperthermophilic crenarchaeon Sulfophobococcus zilligii
- Authors
- Lee, YJ[Lee, YJ]; Choi, JJ[Choi, JJ]; Kwon, ST[Kwon, ST]
- Issue Date
- 1-Apr-2006
- Publisher
- ELSEVIER SCIENCE INC
- Keywords
- DNA polymerase; exonuclease activity; Sulfophobococcus zilligii; archaea; hyperthermophile
- Citation
- ENZYME AND MICROBIAL TECHNOLOGY, v.38, no.6, pp.821 - 830
- Indexed
- SCIE
SCOPUS
- Journal Title
- ENZYME AND MICROBIAL TECHNOLOGY
- Volume
- 38
- Number
- 6
- Start Page
- 821
- End Page
- 830
- URI
- https://scholarworks.bwise.kr/skku/handle/2021.sw.skku/87410
- DOI
- 10.1016/j.enzmictec.2005.08.010
- ISSN
- 0141-0229
- Abstract
- The gene encoding Sulfophobococcus zilligii DNA polymerase (Szi DNA polymerase) was cloned and sequenced. The Szi DNA polymerase gene encompasses 2394 bp, and codes for a protein consisting of 797 amino acid residues. The deduced amino acid sequence of Szi DNA polymerase exhibited a high degree of similarity with archaeal family B-type DNA polymerase homologues found in both crenarchaeotes and euryarchaeotes (Group I), and contained all of the motifs conserved in the homologues for 3' -> 5' exonuclease and polymerase activities. The Szi DNA polymerase gene was expressed under the control of the T7lac promoter on the expression vector pET-28a(+) in Escherichict coli BL21-CodonPlus(DE3)-RIL. The expressed enzyme was purified by heat treatment and Cibacron blue 3GA column chromatography. The optimum pH of the purified enzyme was determined to be in the range of 6.5-7.0. The enzyme was activated by the magnesium ion, and its activity was inhibited by EDTA and monovalent cations, such as potassium ion and ammonium ion. The half-life of the enzyme at 95 degrees C was calculated to be 4.5 h. Szi DNA polymerase possessed associated 3' -> 5' and 5' -> 3' exonuclease activities. (c) 2005 Elsevier Inc. All rights reserved.
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Collections - Biotechnology and Bioengineering > Department of Genetic Engineering > 1. Journal Articles
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