Reference - Detail
| RRC ID | 15988 |
|---|---|
| Author | Baar C, d'Abbadie M, Vaisman A, Arana ME, Hofreiter M, Woodgate R, Kunkel TA, Holliger P. |
| Title | Molecular breeding of polymerases for resistance to environmental inhibitors. |
| Journal | Nucleic Acids Res |
| Abstract |
Potent inhibitors limit the use of PCR assays in a wide spectrum of specimens. Here, we describe the engineering of polymerases with a broad resistance to complex environmental inhibitors using molecular breeding of eight different polymerase orthologues from the genus Thermus and directed evolution by CSR in the presence of inhibitors. Selecting for resistance to the inhibitory effects of Neomylodon bone powder, we isolated 2D9, a chimeric polymerase comprising sequence elements derived from DNA polymerases from Thermus aquaticus, Thermus oshimai, Thermus thermophilus and Thermus brockianus. 2D9 displayed a striking resistance to a broad spectrum of complex inhibitors of highly divergent composition including humic acid, bone dust, coprolite, peat extract, clay-rich soil, cave sediment and tar. The selected polymerase promises to have utility in PCR-based applications in a wide range of fields including palaeobiology, archaeology, conservation biology, forensic and historic medicine. |
| Volume | 39(8) |
| Pages | e51 |
| Published | 2011-4-1 |
| DOI | 10.1093/nar/gkq1360 |
| PII | gkq1360 |
| PMID | 21297114 |
| PMC | PMC3082909 |
| MeSH | Bone and Bones DNA-Directed DNA Polymerase / chemistry DNA-Directed DNA Polymerase / genetics* Directed Molecular Evolution / methods* Enzyme Inhibitors / chemistry Enzyme Inhibitors / pharmacology Gene Library Humic Substances Polymerase Chain Reaction Soil Taq Polymerase Thermus / enzymology |
| IF | 11.502 |
| Times Cited | 46 |
| WOS Category | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| Altmetric score |
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 12 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| General Microbes | JCM 11600 JCM 11601 JCM 11602 JCM 11603 |