Letter abstract
Nature Biotechnology 25, 939 - 943 (2007)
Published online: 15 July 2007 | doi:10.1038/nbt1321
Molecular breeding of polymerases for amplification of ancient DNA
Marc d'Abbadie1, Michael Hofreiter2, Alexandra Vaisman3, David Loakes1, Didier Gasparutto4, Jean Cadet4, Roger Woodgate3, Svante Pääbo2 & Philipp Holliger1
In the absence of repair, lesions accumulate in DNA. Thus, DNA persisting in specimens of paleontological, archaeological or forensic interest is inevitably damaged1. We describe a strategy for the recovery of genetic information from damaged DNA. By molecular breeding2 of polymerase genes from the genus Thermus (Taq (Thermus aquaticus), Tth (Thermus thermophilus) and Tfl (Thermus flavus)) and compartmentalized self-replication3, 4 selection, we have evolved polymerases that can extend single, double and even quadruple mismatches, process non-canonical primer-template duplexes and bypass lesions found in ancient DNA, such as hydantoins and abasic sites. Applied to the PCR amplification of 47,000–60,000-year-old cave bear DNA, these outperformed Taq DNA polymerase by up to 150% and yielded amplification products at sample dilutions at which Taq did not. Our results demonstrate that engineered polymerases can expand the recovery of genetic information from Pleistocene specimens and may benefit genetic analysis in paleontology, archeology and forensic medicine.
- Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK.
- Max Planck Institute for Molecular Anthropology, Department of Evolutionary Genetics, Deutscher Platz 6, D-04103 Leipzig, Germany.
- Section on DNA Replication, Repair and Mutagenesis, Building 6, Room 1A13, National Institutes of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892-2725, USA.
- Laboratoire Lésions des Acides Nucléiques, SCIB/UMR-E 3 (CEA/UJF), Département de Recherche Fondamentale sur la Matière Condensée, Commissariat à l'Énergie Atomique (CEA)/Grenoble, F-38054 Grenoble Cedex 9, France.
Correspondence to: e-mail: ph1@mrc-lmb.cam.ac.uk
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