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Generation of error-prone polymerase variants using directed evolution: Molecular basis of replication fidelity

Fachliche Zuordnung Biochemie
Förderung Förderung von 2004 bis 2011
Projektkennung Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 5427475
 
In the proposed project, the method repertoire of "directed evolution" will be employed for the generation of variants of two viral replication enzymes with decreased fidelity. Polynucleotide polymerases with altered replication error rates are of significant interest for an understanding of the molecular basis of fidelity, and of the natural evolution of polymerase function. The DNA polymerase of bacteriophage T7 and the reverse transcriptase of AIDS virus HIV are targeted in this study because they are well-characterized (biochemically, structurally), and because they differ significantly (factor 60) with respect to their error rates. The results of our study could contribute to an understanding of diverse pathological processes which are related to the fidelity of nucleic acid polymerases. Furthermore, we expect results that are of significant interest for biotechnology because polymerases with altered fidelity or altered substrate tolerance could be employed for the synthesis of nucleic acid libraries, and for diverse amplification processes.
DFG-Verfahren Schwerpunktprogramme
 
 

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