Objective
Research objectives and content The recent genome sequences of several species of archaea indicate the presence of a eukaryotic-type replication 'machinery' in this third domain of life. However, our knowledge about the details of DNA replication in hyperthermophilic archaea is currently limited to the identification of a few putative replication proteins in the genome sequences available, and consequently we propose to initiate a systematic and broad study on mechanistic details of DNA replication in these extremophiles. Using biochemical and molecular biological means, we wish to investigate and identify previously unidentified archaeal replication factors. Their identification should be informative to further our understanding of DNA replication also in eukaryotic systems. We also might be able to identify unique proteins of commercial value that could be used to improve the processivity or fidelity of thermostable DNA polymerases routinely used in PCR applications. Using the plasmid isolated in Dr. Forterre's research group fromPyrococcus abyssii, in vitro assay(s) for hypertherrnophilic primase activity will be established. A yeast two-hybrid system will be used to extensively screen for proteins interacting with the known components of an archaeal replication fork. In addition, in collaboration with Genescope (French National Sequencing center), we will search for additional DNA replication factors in the genome sequence of Pyrococcus abyssii .
Training content (objective, benefit and expected impact)
The host laboratory is an ideal place for the proposed experiments due to the availability of the highly specialised equipment necessary for cultivation of Pyrococcus species as well as the presence of several scientists with the deep knowledge and interest on DNA replication. Optimally, my stay in Orsay will open new doors to studies on DNA replication, and will help to establish myself as an independent scientist. Links with industry / industrial relevance (22)
Dr. Forterre's research group is in contact with European Industries through the Industrial platform of the EC project 'Extremophiles as cell factory'. In addition, we have contacted Finozymes Oy in Espoo, Finland (see the enclosed fax) with the regards of PCR applications for our novel proteins discovered during this project.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- natural sciences biological sciences genetics DNA
- natural sciences biological sciences biochemistry biomolecules proteins
- natural sciences biological sciences genetics genomes
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Coordinator
91405 GOMETZ LA VILLE
France
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