Browsing by Subject "modified base"

Bacteriophages are viruses that infect bacteria. With the ever-increasing threat of antibiotic resistance, they have emerged as a promising alternative treatment. Many phage genomes contain modified bases. They prevent digestion by restriction enzymes allowing the resistance of these viruses to bacterial defence mechanisms. YerA41, a phage that infects Yersinia ruckeri, contains a genome that could not be amplified using any of the DNA polymerases available in the market. Neither restrictions enzymes were able to digest it. These properties led to the assumption that YerA41 genome is not conventional and is likely to contain modified nucleotides. In order to replicate its genome, YerA41 should possess its own DNA polymerase that would be able to use, YerA41 genome as template. If so, it would be able to use other modified genomes as well. Hence, this DNA polymerase could become a very valuable biotechnological tool. In this study we isolated and optimised the purification of DNAP01, one of the putative DNA polymerases encoded by YerA41 genome. In addition, this work shows, with the help of different experiments, how DNAP01 is a novel DNA polymerase able to use YerA41 DNA as template. This is the first time an enzyme of this nature has been described and isolated.