Aim of the project
Do you want to help solve the climate crisis?
Did you know that N2O (laughing gas) has a 300-fold higher impact than CO2 as a greenhouse gas? This means that we need urgent solutions to avoid N2O emissions or to reduce the formed N2O to innocuous nitrogen gas N2.
N2O is formed predominantly because of microbial activity – and microbes can also help us to remove laughing gas (by reducing it to N2) from the atmosphere. The challenge is that N2O respiring bacteria don’t work effectively when O2 is around. Hence, the objective of this project is to engineer recently identified N2O-respiring bacteria, so they become O2-tolerant and O2-indifferent.
You will do this by identifying genes encoding cytochrome c oxidase (responsible for reduction of O2 to water) and deactivating them through selected genome modification strategies (either based on homologue recombination or CRISPR/Cas9 system). Many of the N2O-respiring bacteria harbouring the clade II type nosZ such as strains of the Azospira and Dechloromonas genera are currently not amenable to genetic engineering. This is why the project will first focus on creation of antibiotic selection strategies, (testing of MIC for various antibiotics and the resistance genes), establishment of methodology for cell transformation, plasmid-based gene expression and genome modification. When successful, the newly developed tools will be used to create cytochrome c oxidase-deficient strains.
Methods
Molecular biology (PCR, gel electrophoresis, plasmid isolation, transformation with foreign DNA), basic microbial techniques (media preparation, microbial cultivation, sterile techniques), analytical techniques
This project is a collaboration between the labs of Marta Irla and of Barth F Smets
