Genetic engineering of conjugative relaxases

Lines connect the coordinator with its partners.

Project objective

Research objectives and content In bacterial conjugation, DNA is transferred from donor to recipient bacteria (or to yeast). In T-DNA transfer, structurally and functionally related to conjugation, DNA is transferred from Agrobacterium tumefaciens to plant cells. The aim of the proposed project is to investigate DNA processing during conjugation, using plasmid R388 as a model system. The relaxase protein TrwC is the key enzyme in this process. It is unique among relaxases since it catalyses intermolecular site-specific recombination between two oriT-containing DNA molecules. The biochemistry of TrwC will be analysed in detail, | to convert it to a technological tool. A second unique relaxase is VirD2, the relaxase of Agrobacrerium plasmids, that pilots the T-DNA to the plant cell nucleus. We propose to engineer TrwC by adding some features of VirD2 to achieve high frecuency site-specific DNA integration in yeast, plant and animal cells.Training content (objective, benefit and expected impact) The trainee should become an expert in the biochemistry of relaxases. This technology should have an impact in genetic engineering projects using eukaryotic hosts. Integration of foreign DNA to specific genomic sites will be more efficient and | abolish differences in gene expression and stability common to alternative methods. Other biotechnological applications of these enzymes will be explored.Links with industry / industrial relevance (22) None.

Original text from CORDIS.

Participants

• UNIVERSIDAD DE CANTABRIA · SANTANDERCoordinatorSpain

Links

• View on CORDIS