Consulta: subjectFacets:"DNA polymerase"
Registros recuperados: 9
Data/hora: 06/06/2024 18:20:57

Molecular cloning and expression of key gene encoding hypothetical DNA polymerase from B. mori parvo-like virus
Provedor de dados: 74
Autores: Zhang,Junhong; Li,Guohui; Chen,Huiqing; Li,Xiaogang; Lv,Meng; Chen,Keping; Yao,Qin.
BmPLV-Z is the abbreviation for Bombyx mori parvo-like virus (China isolate). This is a novel virus with two single-stranded linear DNA molecules, viz., VD1 (6543 bp) and VD2 (6022 bp), which are encapsidated respectively into separate virions. Analysis of the deduced amino acid sequence of VD1-ORF4 indicated the existence of a putative DNA-polymerase with exonuclease activity, possibly involved in the replication of BmPLV-Z. In the present study, a recombinant baculovirus was constructed to express the full length of the protein encoded by the VD1-ORF4 gene (3318 bp). In addition, a 2163-bp fragment amplified from the very same gene was cloned into prokaryotic expression vector pET-30a and expressed in E.coli Rosetta 2 (DE3) pLysS. The expressed fusion...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Bombyx mori parvo-like virus; DNA polymerase; Recombinant protein; Sf-9.
Ano: 2010
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572010000400021

La réplication de l'ADN chez l'euryarchaea Pyrococcus Abyssi : mise en place et dynamique du complexe
Provedor de dados: 5
Autores: Rouillon, Christophe.
DNA replication occurs through a large protein complex assembly called replisome. An understanding of its structure and function requires its in vitro reassembly from individual subunits. In all living organisms, the elongation of a primed DNA template by DNA polymerase, with high processivity, requires the accessory protein DNA sliding clamp (PCNA) that is loaded onto DNA by the clamp loader (RF-C). Generally, the archaeal proteins involved in DNA replication are more similar to those of the higher organisms (Eukarya) rather than to those from Bacteria.  This study have focussed on the PCNA loading mechanism and on the DNA synthesis in the hyperthermophilic Euryarchaea Pyrococcus abyssi. To investigate the importance of some protein-protein and...
Tipo: Text
Palavras-chave: Pyrococcus; Archaea; DNA synthesis; Interaction; RP A; RF C; PCNA; DNA polymerase; DNA replication; Pyrococcus; Archée; Archaea; Synthese d'ADN; Interaction; RP A; RF C; PCNA; ADN polymerase; Replication de l'ADN.
Ano: 2006
URL: http://archimer.ifremer.fr/doc/2006/these-2461.pdf

Development of a polymerase chain reaction for the detection of abalone herpesvirus infection based on the DNA polymerase gene
Provedor de dados: 5
Autores: Chen, M.h.; Kuo, S. T.; Renault, Tristan; Friedman, C. S.; Chang, P. H.
A 5781-base pair (bp) fragment of genomic DNA from the Taiwanese abalone herpesvirus was obtained and showed 99% (5767/5779) homology in the nucleotide sequence and 99% (1923/1926) in the amino acid sequence with the DNA polymerase gene of the abalone herpesvirus strain Victoria/AUS/2007. Homology of the amino acid sequence with the DNA polymerase of ostreid herpesvirus 1 was 30% (563/1856). In this study, a PCR-based procedure for detecting herpesvirus infection of abalone, Haliotis diversicolor supertexta, in Taiwan was developed. The method employed primer sets targeting the viral DNA polymerase gene, and was able to amplify DNA fragments of the expected size from infected samples. Primer sets of 40f and 146r were designed for amplification of an...
Tipo: Text
Palavras-chave: Herpes virus; Abalone; Haliotis diversicolor supertexta; DNA polymerase; Polymerase chain reaction.
Ano: 2012
URL: http://archimer.ifremer.fr/doc/00098/20952/19675.pdf

Role of RadA and DNA Polymerases in Recombination-Associated DNA Synthesis in Hyperthermophilic Archaea
Provedor de dados: 5
Autores: Hogrel, Gaelle; Lu, Yang; Alexandre, Nicolas; Bossé, Audrey; Dulermo, Remi; Ishino, Sonoko; Ishino, Yoshizumi; Flament, Didier.
Among the three domains of life, the process of homologous recombination (HR) plays a central role in the repair of double-strand DNA breaks and the restart of stalled replication forks. Curiously, main protein actors involved in the HR process appear to be essential for hyperthermophilic Archaea raising interesting questions about the role of HR in replication and repair strategies of those Archaea living in extreme conditions. One key actor of this process is the recombinase RadA, which allows the homologous strand search and provides a DNA substrate required for following DNA synthesis and restoring genetic information. DNA polymerase operation after the strand exchange step is unclear in Archaea. Working with Pyrococcus abyssi proteins, here we show...
Tipo: Text
Palavras-chave: Homologous recombination; Archaea; DNA polymerase; Recombinase; DNA repair.
Ano: 2020
URL: https://archimer.ifremer.fr/doc/00641/75274/75539.pdf

PCR performance of a thermostable heterodimeric archaeal DNA polymerase
Provedor de dados: 5
Autores: 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

Short Technical Report for OsHV-1 detection and quantification by Real Time Polynerase Chain Reaction using OsHV-1 DNA polymerase sequence
Provedor de dados: 5
Autores: Pepin, Jean-francois.
The present report completes a previously described procedure routinely used for the detection and quantification of OsHV-1 by Sybr green Real Time Polymerase Chain Reaction targeting the OsHV-1 DNA polymerase sequence. More particularly, this document brings information about analytical sensitivity, quantitation limit, assay precision, reproducibility of the technique.
Tipo: Text
Palavras-chave: PCR quantitative HDP-F/R; Virus OsHV-1; Diagnostic; DNA polymerase; Validation; Real-Time PCR HDP-F/R; OsHV-1; Diagnostic; DNA polymerase; Validation.
Ano: 2013
URL: http://archimer.ifremer.fr/doc/00137/24814/22900.pdf

Differential Activities of DNA Polymerases in Processing Ribonucleotides during DNA Synthesis in Archaea
Provedor de dados: 5
Autores: Lemor, Melanie; Kong, Ziqing; Henry, Etienne; Brizard, Raphael; Laurent, Sebastien; Bosse, Audrey; Henneke, Ghislaine.
Consistent with the fact that ribonucleotides (rNTPs) are in excess over deoxyribonucleotides (dNTPs) in vivo, recent findings indicate that replicative DNA polymerases (DNA Pols) are able to insert ribonucleotides (rNMPs) during DNA synthesis, raising crucial questions about the fidelity of DNA replication in both Bacteria and Eukarya. Here, we report that the level of rNTPs is 20-fold higher than that of dNTPs in Pyrococcus abyssi cells. Using dNTP and rNTP concentrations present in vivo, we recorded rNMP incorporation in a template-specific manner during in vitro synthesis, with the family-D DNA Pol (PolD) having the highest propensity compared with the family-B DNA Pol and the p41/p46 complex. We also showed that ribonucleotides accumulate at a...
Tipo: Text
Palavras-chave: Archaea; DNA replication and repair; DNA polymerase; Nucleotide pool; Translesion synthesis.
Ano: 2018
URL: https://archimer.ifremer.fr/doc/00464/57603/59796.pdf

The hyperthermophilic euryarchaeota Pyrococcus abyssi likely requires the two DNA polymerases D and B for DNA replication
Provedor de dados: 5
Autores: Henneke, Ghislaine; Flament, Didier; Hubscher, Ulrich; Querellou, Joel; Raffin, Jean-paul.
DNA polymerases carry out DNA synthesis during DNA replication, DNA recombination and DNA repair. During the past five years, the number of DNA polymerases in both eukarya and bacteria has increased to at least 19 and multiple biological roles have been assigned to many DNA polymerases. Archaea, the third domain of life, on the other hand, have only a subset of the eukaryotic-like DNA polymerases. The diversity among the archaeal DNA polymerases poses the intriguing question of their functional tasks. Here, we focus on the two identified DNA polymerases, the family B DNA polymerase B (PabpolB) and the family D DNA polymerase D (PabpolD) from the hyperthermophilic euryarchaeota Pyrococcus abyssi. Our data can be summarized as follows: (i) both Pabpols are...
Tipo: Text
Palavras-chave: DNA polymerase; Strand displacement; Gap filling; Euryarchaea; DNA replication.
Ano: 2005
URL: http://archimer.ifremer.fr/doc/2005/publication-423.pdf

In vitro reconstitution of RNA primer removal in Archaea reveals the existence of two pathways
Provedor de dados: 5
Autores: Henneke, Ghislaine.
Using model DNA substrates and purified recombinant proteins from Pyrococcus abyssi. I have reconstituted the enzymatic reactions involved in RNA primer elimination in vitro. In my dual-labelled system, polymerase D performed efficient strand displacement DNA synthesis, generating 5'-RNA flaps which were subsequently released by Fen1, before ligation by Lig1. In this pathway, the initial cleavage event by RNase HII facilitated RNA primer removal of Okazaki fragments. In addition, I have shown that polymerase B was able to displace downstream DNA strands with a single ribonucleotide at the 5'-end, a product resulting from a single cut in the RNA initiator by RNase HII. After RNA elimination, the combined activities of strand displacement DNA synthesis by...
Tipo: Text
Palavras-chave: Archaea; DNA ligase; DNA polymerase; Nuclease; Okazaki fragment.
Ano: 2012
URL: http://archimer.ifremer.fr/doc/00107/21841/20923.pdf