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| Killelea, Tom; Ralec, Celine; Bosse, Audrey; Henneke, Ghislaine. |
| DNA polymerases are versatile tools used in numerous important molecular biological core technologies like the ubiquitous polymerase chain reaction (PCR), cDNA cloning, genome sequencing, and nucleic acid based diagnostics. Taking into account the multiple DNA amplification techniques in use, different DNA polymerases must be optimized for each type of application. One of the current tendencies is to reengineer or to discover new DNA polymerases with increased performance and broadened substrate spectra. At present, there is a great demand for such enzymes in applications, e.g., forensics or paleogenomics. Current major limitations hinge on the inability of conventional PCR enzymes, such as Taq, to amplify degraded or low amounts of template DNA. Besides,... |
| Tipo: Text |
Palavras-chave: DNA polymerase; Archaea; Family D; PCR; Pyrococcus. |
| Ano: 2014 |
URL: http://archimer.ifremer.fr/doc/00193/30450/28871.pdf |
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| Ralec, Celine; Henry, Etienne; Lemor, Melanie; Killelea, Tom; Henneke, Ghislaine. |
| Divalent metal ions, usually Mg2+, are required for both DNA synthesis and proofreading functions by DNA polymerases (DNA Pol). Although used as a non-reactive cofactor substitute for binding and crystallographic studies, Ca2+ supports DNA polymerization by only one DNA Pol, Dpo4. Here, we explore whether Ca2+-driven catalysis might apply to high-fidelity (HiFi) family B DNA Pols. The consequences of replacing Mg2+ by Ca2+ on base pairing at the polymerase active site as well as the editing of terminal nucleotides at the exonuclease active site of the archaeal Pyrococcus abyssi DNA Pol (PabPolB) are characterized and compared to other (families B, A, Y, X, D) DNA Pols. Based on primer extension assays, steady-state kinetics and ion-chased experiments, we... |
| Tipo: Text |
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| Ano: 2017 |
URL: http://archimer.ifremer.fr/doc/00405/51663/52210.pdf |
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| Ralec, Celine. |
| Hyperthermophilic Archaea that thrive under harsh environments (elevated temperature, pH shifts, andionizing radiations) are supposed to be exposed to massive DNA damages. However, the mutations frequencies in hyperthermophilic Archaea are comparable with those of other microorganisms (1) indicating they are equipped with unique and efficient molecular mechanisms to ensure their genome integrity. DNA replication is an essential and conserved process among the three domains of life. DNA polymerases are central enzymes involved in the joining of deoxyribonucleoside 5′-triphosphates (dNTPs) to form the growing DNA chain. In Pyrococcus abyssi (Pab), two familiesDNA polymerases have been described as replicases, one family B (PabPol B) with structural diversity... |
| Tipo: Text |
Palavras-chave: ADN; Archéa; Polymérase; Pyrococcus. |
| Ano: 2013 |
URL: https://archimer.ifremer.fr/doc/00498/60952/64355.pdf |
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