Effect of endogenous nucleases on natural transformation of Actinetobacter baylyi ADP1

Acinetobacter baylyi ADP1 is a gram-negative bacterium with the ability to uptake extracellular DNA and integrate it into its genome. While ADP1’s natural transformation has been studied with respect to homologous regions and DNA damage, the effect of nucleases on this ability is not known. Our hypothesis is that the transformation event is a race between integration and degradation or fragmentation of internalized DNA. By creating nuclease knockout strains we can characterize their impact on transformation frequency, and hopefully increase the viability of ADP1 as a model organism for large-scale genome engineering. Through studying the three knockout strains successfully created we have found no statistically significant change in transformation efficiency, and are expanding our analysis to a greater breadth of nucleases.