Purification and characterization of Thermus caldophilus GK24 DNA polymerase

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Title
Purification and characterization of Thermus caldophilus GK24 DNA polymerase
Author(s)
Jong Hoon Park; Joon Su Kim; Suk Tae Kwon; Dae Sil Lee
Bibliographic Citation
European Journal of Biochemistry, vol. 214, pp. 135-140
Publication Year
1993
Abstract
A thermostable DNA polymerase from Thermus caldophilus GK24 was purified to near homogeneity by chromatographic methods, including ion-exchange, gel-filtration and affinity chromatography. The purified enzyme had a specific activity of 8400 U/mg at 75°C and a lecular mass of 95 kDa, estimated by SDS/PAGE and Superose-12 gel filtration. Reaction conditions were investigated in terms of pH, metal-ion concentration and temperature. Experimental results showed that T. caldophilus (Tca) DNA polymerase had a maximum activity near pH 8.7 at 75 °C. The N-terminal sequence of the enzyme was highly similar to that of Thermus aquaticus (Taq) DNA polymerase, which was consistent with the fact that the enzyme had 5'-to-3' exonuclease activity and no 3'-to-5' exonuclease activity. Gene amplification using Tca DNA polymerase resulted in longer products than amplification using Taq DNA polymerase.
Keyword
dna polymeraseenzyme purificationthermusDNA-directed DNA polymerasepolymerase chain reaction
ISSN
0014-2956
Publisher
Wiley
DOI
http://dx.doi.org/10.1111/j.1432-1033.1993.tb17905.x
Type
Article
Appears in Collections:
Division of Bio Technology Innovation > SME Support Center > 1. Journal Articles
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