Project Details
Projekt
Generation of error-prone polymerase variants using directed evolution: Molecular basis of replication fidelity
Applicant
Professorin Dr. Susanne Brakmann
Subject Area
Biochemistry
Term
from 2004 to 2011
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 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 Programme
Priority Programmes
