Plant Secondary Metabolism Engineering

This book presents detailed practical information on important methods used in the engineering of plant secondary metabolism pathways and the acquisition of essential knowledge in performing this activity, including important advances and emerging strategies.

Plants have evolved an amazing array of metabolic pathways leading to molecules capable of responding promptly and effectively to stress situations imposed by biotic and abiotic factors, some of which supply the ever-growing needs of humankind for natural chemicals, such as pharmaceuticals, nutraceuticals, agrochemicals, food and chemical additives, biofuels, and biomass. In Plant Secondary Metabolism Engineering: Methods and Applications , expert researchers provide detailed practical information on some of the most important methods employed in the engineering of plant secondary metabolism pathways and in the acquisition of essential knowledge in performing this activity, including the significant advances and emerging strategies. 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 notes on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Plant Secondary Metabolism Engineering: Methods and Applications will aid scientists engaged in the challenging task of modifying some of the most intricate products of plant evolution and support their efforts directed toward the vital goal of sustainable natural chemicals.

Provides an easily accessible reference source of strategies for engineering plant secondary metabolic pathways Details the methods to introduce, manipulate, up and down regulate, and evaluate the expression of plant metabolism genes in both homologous and heterologous systems Supplies practical knowledge toestablish viable systems for the demand of natural products as pharmaceuticals, food additives, nutraceuticals, and special chemicals Includes supplementary material: sn.pub/extras

Inhalt
Plant Secondary Metabolism and Challenges in Modifying Its Operation: An Overview.- Suppression Subtractive Hybridization as a Tool to Identify Anthocyanin Metabolism-Related Genes in Apple Skin.- Identification of Regulatory Protein Genes Involved in Alkaloid Biosynthesis Using a Transient RNAi System.- Site-Directed Mutagenesis and Saturation Mutagenesis for the Functional Study of Transcription Factors Involved in Plant Secondary Metabolite Biosynthesis.- Isolation of Proteins Binding to Promoter Elements of Alkaloid Metabolism-Related Genes Using Yeast One-Hybrid.- Modulation of Carotenoid Accumulation in Transgenic Potato by Inducing Chromoplast Formation with Enhanced Sink Strength.- Over-expression of Rate-Limiting Enzymes to Improve Alkaloid Productivity.- Microbial Expression of Alkaloid Biosynthetic Enzymes for Characterization of Their Properties.- Producing a Recombinant Flavin-Containing Monooxygenase from Coffea arabica in Escherichia coli for Screening of Potential Natural Substrates.- Efficient Production of Active Form Recombinant Cassava Hydroxynitrile Lyase Using Escherichia coli in Low-Temperature Culture.- of the Early Pathway to Taxol Biosynthesis in Yeast by Means of Biosynthetic Gene Cluster Construction Using SOE-PCR and Homologous Recombination.- Biocatalytic Synthesis of Tritium (3H)-Labelled Taxa-4(5),11(12)-diene, the Pathway Committing Precursor of the Taxoid Diterpenoids.- USER Cloning and USER Fusion: The Ideal Cloning Techniques for Small and Big Laboratories.- Enrichment of Carotenoids in Flaxseed by Introducing a Bacterial Phytoene Synthase Gene.- Metabolic Engineering by Plastid Transformation as a Strategy to Modulate Isoprenoid Yield in Plants.- Engineering High Yields of Secondary Metabolites in Rubia Cell Cultures Through Transformation with Rol Genes.- Flow Cytometric Methods to Investigate Culture Heterogeneities for Plant Metabolic Engineering.- Phenylpropanoid Biosynthesis in Leaves and Glandular Trichomes of Basil (Ocimum basilicum L.).- Fusion with Fluorescent Proteins for Subcellular Localization of Enzymes Involved in Plant Alkaloid Biosynthesis.- Immunohistochemical Localisation of a Putative Flavonoid Transporter in Grape Berries.- Electrogenic Bromosulfalein Transport in Isolated Membrane Vesicles: Implementation in Both Animal and Plant Preparations for the Study of Flavonoid Transporters.