We would like to use C. elegans as a biosensor for environmental toxins. We are investigating the feasibility of transforming C. elegans with luminescence reporter genes placed under constitutive promoters and of using light output to measure its metabolic status. We should then be able to assess how various xenobiotics affect C. elegans metabolism as measured by light output. Conversely, we should be able to detect toxins by their effect on bioluminescence. The most frequently used luminescence reporter genes are: the luc gene, cloned from the firefly Photinus pyralis, and the lux genes, from the marine bacteria Vibrio harveyi and Vibrio fisheri. The luc gene encodes an enzyme, firefly luciferase, which catalyses the oxidation of luciferin in a reaction that consumes ATP and produces light. Therefore, when the gene is expressed constitutively and luciferin is readily available, the light output is correlated to cellular ATP pools. The luxAB genes encode a bacterial luciferase which catalyses the oxidation of FMNH2\/NADH and a long-chain aliphatic aldehyde (e.g. n-decylaldehyde) resulting in light emission. Using these approaches metabolism can be linked to bioluminescence through the FMNH2\/NADH pools. Both luciferin and n-decylaldehyde have to be provided exogenously to the nematodes. Xenobiotics will disrupt the metabolism, leading to a decrease in the cellular energy reserves and consequently to a decrease in bioluminescence. Low concentrations of n-decylaldehyde proved to be toxic to C. elegans, hence we concentrated on the luc gene. This gene was fused to a let-858 promoter and injected into wildtype animals together with the pRF4 plasmid. Permeability of the substrate (luciferin) might pose a challenge to the development of bioluminescent nematodes. Once these are obtained , we will quantify their response to concentration ranges of different environmental toxins.
|Publication status||Published - 1998|
|Event||European Worm Meeting - Hinxton, Cambridge, United Kingdom|
Duration: 19 Feb 2018 → …
|Conference||European Worm Meeting|
|Period||19/02/18 → …|