Project Description
Oil sands operations in Northern Alberta bring economic prosperity to Canada. But, they also produce contaminated air, water and soil. We are in the process of transforming the nematode worm Caenorhabditis elegans into a soil bioremediation toolkit with a specific focus on naphthalene.
Wild-type C. elegans have a weak chemotaxis response to naphthalene. However, we will enhance this chemotaxis response using G-protein coupled receptors (GPCRs) found in H. sapiens, M. musculus, and R. norvegicus. Our goal is to import these foreign GPCRs into the worm under control of C. elegans neuron-specific promoters. This should produce a transgenic worm with enhanced capacity to seek out naphthalene.
We will then design a field bioassay based on our C. elegans chemotaxis system. We envision an agar plate with two populations of worms expressing different types of fluorescent protein. One population will chemotax toward naphthalene, and the other will be repulsed by naphthalene.The presence of naphthalene in a soil sample in the middle of the plate would be indicated by concentric rings of fluorescence forming around the sample.
We will also equip C. elegans with the capacity to degrade naphthalene. The NAH7 plasmid in Pseudomonas putida encodes all of the enzymes necessary break naphthalene down to pyruvate. Our ultimate goal is to bring each NAH7 gene into C. elegans under control of one of the worm’s constitutive promoters. For our project this summer, we will focus on the first enzyme in the pathway, encoded by the gene nahA. If we can demonstrate the working of this enzyme in C. elegans cells, it will provide a proof of concept that encourages further work on this pathway.
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