The main objective is testing how important the RecFOR pathway genes RecO and RecR are in Thermus thermophilus biology, toghether with the downstream resolvase RuvB, whose product is part of the RuvABC complex.

Brief description of the experiment: DNA repair pathways are important in the biology of the bacterium Thermus thermophilus. We have observed that genome repair by homologous recombination is very active in this organism and, probably, and essential process. In the present work we are interested in testing how important the RecFOR pathway genes RecO and RecR are in Thermus thermophilus biology, toghether with the downstream resolvase RuvB, whose product is part of the RuvABC complex. RecO and RecR deletion mutants present clear defects in growth and transformation while RuvB grows normally but is unable to integrate a linear DNA that has to undergone double recombination for its integration in the genome. Previous evidence suggests that the deletion or inactivation of the genes AddA and/or AddB rescues the defective growth and transformation phenotype of mutants in the RecFOR pathway. In this sequencing project we want to check the genotype of hygromycin insertion mutants in RecO, RecR or RuvB (3 strains of each mutant) in a clean AddAB deletion strain background of Thermus thermophilus. Plus, another three RuvB mutants in the wild-type HB27 (ATCC BAA- 163/DSM7039) background. The idea is to detect possible compensatory mutations that could have appeared in the mutant strains to alleviate the growth defects observed in them. As controls, we sequenced the clean ?AddAB strain and that strain with a hygromycin resistance gene inserted at the TT_C0313 locus. The objective of this project is to analyse the Single Nucleotide Polymorphisms (SNPs) between mutants and its parental strains, and check whether the mutants genotype is what is expected.