Investigating oxidative stress and large-scale genomic reduction in Vibrio natriegens through the development and application of new genetic techniques

Vibrio natriegens is the fastest replicating bacterium and recent work has demonstrated its potential for biotechnology and synthetic biology applications. While these industries have been the focus of V. natriegens research, it offers a promising system for diverse laboratory studies. However, currently a complete understanding of V. natriegens biology is lacking. As such, there is a need to advance genetic techniques and physiological understanding to better inform all aforementioned applications. Here we describe our contributions to the growing field of V. natriegens genetics and the application of these new techniques to study physiological processes and themes in genome evolution. Briefly, we performed a case study on a set of physiologically important genes natively found in V. natriegens genome, called katG1, katG2, and katG3, whose function is to protect the cell from oxidative stress. We report functional and phylogenetic analyses that inform the evolutionary and ecological basis for katG multiplicity in V. natriegens. We next expand our analysis to investigate generalized genome reduction as a simulation of genome streamlining. Genome streamlining theory posits that reduction of DNA content is selectively advantageous under persistent nutrient limitation. V. natriegens has an expansive genome and we leverage this characteristic and apply our new genetic techniques to investigate the physiological impacts of large-scale genomic reductions under theoretically relevant conditions. Together the work presented here expands the capabilities and understanding of V. natriegens has a host for molecular biology.