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The use of DNA repair as a tool in the study of factors involved in pathogenicity of Corynebacterium genus and characterization of a DNA repair gene present in pathogenic species.

The DNA repair pathway is an important mechanism in the maintenance of the genomic stability and the vulnerability of this system makes it an interesting target in search on the control of infectious organisms. The goal of this work was to identify and characterize DNA repair pathways of eight species of Corynebacterium, pathogenic or not, and select and characterize an enzyme, present only in pathogenic species, through in vitro assays to confirm its involvement in DNA repair. Was verified that the pathways of DNA repair in general are well conserved and NER and recombination pathways showed having virtually all conserved genes. The enzyme selected, uracil DNA glycosilase, is important to remove uracil from DNA with can cause transversion C/G into T/A during a replication event. The mug gene sequence of C.pseudotuberculosis, obtained from CoryneRegNet databank, was amplified by PCR and cloned in expression vector pET21 to be sequenced. To confirm expression in E. coli BL21 strain was carried out a western blot assay. The protein was purified and is currently being tested in in vitro experiments to verify its ability to recognize uracil in oligos sintehtyzed. An understanding of the precise molecular functions of the enzymes participating in DNA metabolism and in the maintenance of pathogen genome integrity can be the key to search for targets to be used in therapeutic intervention. Financial support: FAPEMIG