Molecular and Cellular Bioengineering Research Experiences for Undergraduates (REU) Program
Making maps of carbon traffic in plant cells through isotope labeling-assisted flux quantification
Metabolic pathways are sequences of biochemical reactions in cells, and our lab focuses on quantifying carbon traffic ("flux") through networks of interconnected metabolic pathways. This is relevant because information from flux analysis is crucial to understanding plant cell function and in identifying metabolic engineering strategies, and not much is currently known about regulation of carbon flux in plants. This project will involve investigating cells and tissues of the model plant Arabidopsis with regard to how different nutrients can affect carbon fluxes in this plant. To measure carbon fluxes, we will culture the cells on mixtures of different carbon isotopes (e.g. ^13 C and ^12 C), and then measure isotope labeling patterns in the cells by gas chromatography-mass spectrometry (GCMS) and/or two-dimensional nuclear magnetic resonance (2-D NMR). The isotope labeling patterns thus generated will be processed computationally to evaluate carbon fluxes and make maps of carbon traffic in the cells. Thus, this project will provide exposure to cell culture, analytical chemistry and spectroscopy techniques, as well as computational techniques to analyze metabolic networks. The outcomes of this project—flux maps from plants or plant cells grown under different nutrient conditions—are expected to reveal metabolic nodes or intersections that restrict carbon traffic and thus form bottlenecks. Thus, this project will provide insights toward smartly identifying genetic engineering targets for the plant and has implications on metabolic engineering of plant cells for food and fuel production.
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