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Purification and Characterization of the N-terminally Truncated DNA Polymerase from Thermus thermophilus HJ6  

Jeon, Sung-Jong (Department of Biotechnology & Bioengineering, Dong-Eui University)
Seo, Min-Ho (Department of Biotechnology & Bioengineering, Dong-Eui University)
Publication Information
Microbiology and Biotechnology Letters / v.38, no.2, 2010 , pp. 158-162 More about this Journal
Abstract
The gene encoding N-terminally truncated Tod polymerase ($\Delta$Tod polymerase) from Thermus thermophilus HJ6 was expressed in Escherichia coli under the control of the lambda pR and pL tandem promoters on the expression vector pJLA503. The N-terminal domain (250 amino acids) of Tod polymerase was removed without significant effect on enzyme activity and stability, while no 5'$\rightarrow$3' exonuclease activity was detected. The $\Delta$Tod polymerase was verified to possess very efficient reverse transcriptase (RT) activity in the presence of $MgCl_2$. The cDNA can also be amplified in the polymerase chain reaction (PCR) with this mutant enzyme. The $\Delta$Tod polymerase was exhibited higher activity than the Taq polymerase in a one-step RT-PCR.
Keywords
DNA polymerase; RT-PCR; Thermus thermophilus; PCR; exonuclease;
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