Mechanisms of mutation in Escherichia coli: analysis, engineering and applications

Informations

Funding country

Hungary

Acronym

-

URL

-

Start date

4/1/2008

End date

3/31/2011

Budget

71,133 EUR

Fundings

Abstract

In this study, we plan to investigate the factors influencing the rate and control of mutations in the genome of Escherichia coli. A unique tool, E. coli strain MDS42, engineered in our laboratory is readily available for this purpose. This strain lacks all mobile genetic elements, and thus allows the tight control of the type and copy number of such elements reintroduced on plasmids or on the genome. We also wish to study the changes in the mutation rate after deleting the genes of error-prone DNA-polymerases. Finally, we will attempt to decrease the rate of deletions forming in the genome and on plasmids by overexpressing the genes of certain exonucleases. The combinations of the above mentioned modifications will be tested in order to develop a strain with a lower overall mutation rate and thereby higher genetic stability. This strain could prove to be advantageous in DNA-cloning and protein overproduction purposes compared to present day host strains, and could serve as their replacement within a time scale of 2-5 years. Preliminary experiments conducted in this area have already yielded improved laboratory strains. Improving the understanding of basic mutational mechanisms can have practical benefits in applications related to health and biotech industry as well. This grant would widen the class of young investigators between the age of 30 and 40 in the BRC. Two PhD students, working on related topics under the supervision of my main collaborator, György Pósfai, would indirectly also benefit from the grant. Our working conditions will be improved by the installation of a polyacrlylamide-gel electrophoresis system equipped with blotting module, and a personal computer.