| Title |
Biochemical Properties and PCR Performance of a Family B DNA Polymerase from Hyperthermophilic Euryarchaeon Thermococcus peptonophilus
|
|---|---|
| Published in |
Applied Biochemistry and Biotechnology, May 2009
|
| DOI | 10.1007/s12010-009-8658-0 |
| Pubmed ID | |
| Authors |
Jong Il Lee, Yun Jae Kim, Heejin Bae, Sung Suk Cho, Jung-Hyun Lee, Suk-Tae Kwon |
| Abstract |
The Thermococcus peptonophilus (Tpe) DNA polymerase gene was expressed under the control of the T7lac promoter on pET-22b(+) in Escherichia coli BL21-CodonPlus(DE3)-RIL in order to fully elucidate its biochemical properties and evaluate its feasibility in polymerase chain reaction (PCR) application. The expressed enzyme was then purified by heat treatment followed by two steps of column chromatography after which optimum pH and temperature of the enzyme were evaluated to be 7.0 and 75 degrees C, respectively. The optimal buffer for PCR with Tpe DNA polymerase consisted of 50 mM Tris-HCl (pH 8.0), 2 mM MgCl(2), 80 mM KCl, and 0.02% Triton X-100. Tpe DNA polymerase revealed a 3.6-fold higher fidelity (3.37 x 10(-6)) than Taq DNA polymerase (12.13 x 10(-6)) and performed significantly more efficiently in PCR amplification than both Taq and Pfu DNA polymerases. Ratios of 31:1 of Taq to Tpe DNA polymerases allowed PCR amplification of targets up to 15 kb in length with a 2.2-fold higher fidelity than Taq DNA polymerase. The results of the PCR experiments indicate that Tpe DNA polymerase may provide a higher fidelity DNA amplification in a shorter reaction time. |
Mendeley readers
The data shown below were compiled from readership statistics for 26 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Sweden | 1 | 4% |
| Germany | 1 | 4% |
| Unknown | 24 | 92% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Master | 6 | 23% |
| Researcher | 5 | 19% |
| Student > Bachelor | 4 | 15% |
| Professor | 3 | 12% |
| Student > Ph. D. Student | 2 | 8% |
| Other | 2 | 8% |
| Unknown | 4 | 15% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 11 | 42% |
| Biochemistry, Genetics and Molecular Biology | 6 | 23% |
| Nursing and Health Professions | 1 | 4% |
| Computer Science | 1 | 4% |
| Immunology and Microbiology | 1 | 4% |
| Other | 2 | 8% |
| Unknown | 4 | 15% |
Attention Score in Context
This research output has an Altmetric Attention Score of 3. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 01 October 2016.
All research outputs
#7,454,951
of 22,790,780 outputs
Outputs from Applied Biochemistry and Biotechnology
#541
of 2,503 outputs
Outputs of similar age
#32,576
of 92,751 outputs
Outputs of similar age from Applied Biochemistry and Biotechnology
#10
of 24 outputs
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So far Altmetric has tracked 2,503 research outputs from this source. They receive a mean Attention Score of 3.2. This one is in the 43rd percentile – i.e., 43% of its peers scored the same or lower than it.
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We're also able to compare this research output to 24 others from the same source and published within six weeks on either side of this one. This one is in the 8th percentile – i.e., 8% of its contemporaries scored the same or lower than it.