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Full Description
In Plant Metabolic Flux Analysis, expert researchers in the field provide detailed experimental procedures for each step of the flux quantification workflow. Steady state and dynamic modeling are considered, as well as recent developments for the reconstruction of metabolic networks and for a predictive modeling. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and key tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical Plant Metabolic Flux Analysis, seeks to aid scientists in the further study of cutting-edge protocols and methodologies that are crucial to getting ahead in MFA.
Contents
Application of Metabolic Flux Analysis to Plants.- Metabolic Network Reconstruction and Their Topological Analysis.- 14C pulse Labeling to Estimate External Fluxes and Turnovers in Primary Metabolism.- Optimization of Steady-state 13C-labeling Experiments for Metabolic Flux Analysis.- Quantification of 13C Enrichments and Isotopomer Abundances for Metabolic Flux analysis using 1D NMR Spectroscopy.- Analysis of Proteinogenic Amino Acid and Starch Labeling by 2D NMR.- Analysis of Kinetic Labeling of Amino Acids and Organic Acids by GC-MS.- Quantifying 13C-labeling in Free Sugars and Starch by GC-MS.- Liquid Chromatography Tandem Mass Spectrometry for Measuring 13C-labeling in Intermediates of the Glycolysis and Pentose-phosphate Pathway.- In Vivo NMR for 13C metabolic Flux Analysis.- Steady State and Instationary Modeling of Proteinogenic and Free Amino Acid Isotopomers for Flux Quantification.- Isotopically Nonstationary MFA (INST-MFA) of Autotrophic Metabolism.- Simulating Labeling to Estimate Kinetic Parameters for Flux Control Analysis.- High-throughput Data Pipelines for Metabolic Flux Analysis in Plants.- Analysis of Enzyme Activities.- Analytical Kinetic Modeling: A Practical Procedure.- Flux Balance Analysis as an Alternative Method to Estimate Fluxes Without Labeling.- Flux Variability Analysis: Application to Developing Oilseed Rape Embryos using Toolboxes for Constraint-Based Modeling.- Plant Genome-Scale Modeling and Implementation.- 34S and 15N Labelling to Model S and N Flux in Plants and Determine the Different Components of N and S use Efficiency.- Ecophysiological Process-based Model to Simulate Carbon Fluxes in Plants.
