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[Recombinant Thermus thermophilus His6-DNA polymerase with reverse transcriptase activity]
Bioorganicheskaia Khimiia
|April 1, 1997
Summary
Researchers cloned Thermus thermophilus DNA polymerase genes into E. coli. The purified enzyme shows strong DNA polymerase and reverse transcriptase activity, useful for molecular biology.
Area of Science:
- Molecular Biology
- Biochemistry
- Genetics
Context:
- Thermus thermophilus is a thermophilic bacterium known for its robust enzymes.
- DNA polymerases are crucial enzymes for DNA replication and repair.
- Reverse transcriptases are essential for converting RNA into DNA.
Purpose:
- To clone and express the Tth DNA polymerase genes from Thermus thermophilus.
- To isolate and characterize the Tth DNA polymerase enzyme.
- To assess the enzymatic activities of the purified Tth DNA polymerase.
Summary:
- The Tth DNA polymerase genes from Thermus thermophilus strains HB-8 and Tt-111 were successfully cloned and expressed in Escherichia coli.
- A polyhistidine tag facilitated single-stage purification of the target protein via affinity chromatography on Ni-NTA-agarose.
- The isolated Tth DNA polymerase exhibited significant DNA polymerase and reverse transcriptase activities.
Impact:
- Provides a method for producing active Tth DNA polymerase in a heterologous host.
- The enzyme's dual activity makes it a valuable tool for various molecular biology applications.
- Facilitates further studies on the structure and function of thermophilic DNA polymerases.