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Cloning and Expression of the Gene Encoding DNA Polymerase from Thermus thermophilus HJ6 in Escherichia coli

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The 49th International Symposium of Korean Society of Life Science 37

P19

Cloning and Expression of the Gene Encoding DNA Polymerase from

Thermus thermophilus HJ6 in Escherichia coli

Min-Ho Seo1, Bu-Kyoung Kim1, Pyung-Hwa Kwak1 and Sung-Jong Jeon1,2* 1

Department of Biotechnology & Bioengineering, 2Department of Biomaterial Control (BK21 program), Dong-Eui University, Busan 614-714, Korea

A gene encoding for a putative DNA polymerase (Tod) from Thermus thermophilus HJ6 was cloned and expressed in Escherihcia coli. Tod gene is composed of 2,505 nucleotides, encoding a protein (834 amino acids) with a predicted molecular mass of 93,795 Da from T. thermophilus HJ6. The gene from T. thermophilus encoding a 93.8-kDa protein showed a 79-98% identity with other DNA polymerases from Thermus strains. Tod gene was expressed in E. coli codon Plus (DE3) cells by heat shock induction using the pJLA503 vector system. The E. coli cells were disrupted by sonication and the supernatant fraction was heat-treated at 85oC for 20min. The recombinant Tod has a molecular mass of 94 KDa, determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. DNA amplification using the recombi-nant Tod DNA polymerase performed the same as other thermostable DNA polymerase from Thermus strains. [This work was supported by the Marine and Extreme Genome Research Center Program of the Ministry of Land, Transportation and Maritime Affairs, Republic of Korea]

Key words: Thermus thermophilus, DNA polymerase, heat shock, Thermus, PCR

P20

Effects of Omija and Maesil Extracts in Ionic Water on Endurance

Exercise Performance in Mice

Mi Hee Woo1, Moch. Saiful Bachri2, Jong Won Choi2,3 and Hee Sook Kim1,3 1

Department of Food Science and Biotechnology, Kyungsung University

2

Department of Pharmacology, Kyungsung University

3

Biotech Institute of Kyungsung University

We investigated the effect of new sports drinks, containing 6% carbohydrate, electrolytes, Schizandra chinesis, Prunus

mume and molasses, on the endurance exercise capacity and the changes of blood biochemical parameter in experimental

animals orally administered for 4 weeks. Drinks containing 6% carbohydrate-electrolyte like commercial ionized drinks did not affect to enhance the forced swimming capacity or running time on a treadmill until all-out state. Whereas the addition of Omija (Schisandra berry of Schizandra chinesis) and/or Maesil (fruit of Prunus mume) in 6% carbohy-drate-eletrolyte drinks exhibited increasing effect on the exercise endurance in mice. The addition of both Omija and

Maesil in ionic water showed synergistic effects significantly on the exercise capacity and decreasing effect on the

level of blood lactate and ammonia, metabolites known to cause to fatigue more than the addition of only Omija or Maesil in ionic water. Lignans including Schisandrin from Omija is known as an antihepatotoxic, antiasthmatic, antidiabetic, sedative and tonic agent. Maesil, which has abundant organic acids (succinic acid, citric acid, malic acid and tartaric acid), sitosterol, and minerals, has been commonly used in folk remedies and Chinese medicine over very long period. Acknowledgement: This work was supported by the 2008 Busan TechnoPark program (BTP) and Human Resource Training Project for Strategic Technology.

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