This volume explores plastid evolution, structure, and function in algae, plants and protists. The methods described in this book help scientists visualize, fractionate, purify, and study primary and secondary plastids in plant and algal materials. The chapters in this book also look at various techniques to analyze plastids through means of combining biology strategies from genetics, genomics, proteomics, and lipidomics. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and…mehr
This volume explores plastid evolution, structure, and function in algae, plants and protists. The methods described in this book help scientists visualize, fractionate, purify, and study primary and secondary plastids in plant and algal materials. The chapters in this book also look at various techniques to analyze plastids through means of combining biology strategies from genetics, genomics, proteomics, and lipidomics. 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 tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and thorough, Plastids: Methods and Protocols is a valuable resource for students, engineers, and researchers who are interested in this evolving organelle and overall field.
Primary Endosymbioses: Emergence of the Primary Chloroplast and the Chromatophore, Two Independent Events.- Complex Endosymbioses I: From Primary to Complex Plastids, Multiple Independent Events.- Complex Endosymbiosis II: The Non-Photosynthetic Plastid of Apicomplexa Parasites (The Apicoplast) and Its Integrated Metabolism.- Diversity and Plasticity of Plastids in Land Plants.- The Main Functions of Plastids.- Plastid Transient and Stable Interactions with Other Cell Compartments.- Imaging Plastids in 2D and 3D: Confocal and Electron Microscopy.- Purification of Chloroplasts and Chloroplast Subfractions: Envelope, Thylakoids, and Stroma - From Spinach, Pea, and Arabidopsis thaliana.- Isolation of Inner and Outer Membranes of the Chloroplast Envelope from Spinach and Pea.- Purification of Non-Green Plastids (Proplastids and Amyloplasts) from Angiosperms, and Isolation of Their Envelope Membranes.- In Vitro Protein Import into Isolated Chloroplasts.- Analysis of the MTL Super-Complex at Contact Sites between Mitochondria and Plastids.- Isolation of Plastid-Fractions from the Diatoms Thalassiosira pseudonana and Phaeodactylum tricornutum.- Isolating the Plasmodium falciparum Apicoplast using Magnetic Beads.- Extraction and Quantification of Lipids from Plant or Algae.- Quantitative Assessment of the Chloroplast Lipidome.- Determination of the DNA/RNA-Associated Sub-Proteome from Chloroplasts and Other Plastid Types.- Mapping Plastid Transcript Population by Circular Reverse Transcription-Polymerase Chain Reaction.- Bioinformatic Analysis of Chloroplast Gene Expression and RNA Post-Transcriptional Maturations using RNA Sequencing.- A Guide to the Chloroplast Transcriptome Analysis using RNA-Seq.- A Toolkit for the Characterization of the Photoprotective Capacity of Green Algae.- Rescue of Deletion Mutants to Isolate Plastid Transformants in Higher Plants.- Genetic Analysis of Chloroplast Biogenesis, and Function and Mutant Collections.- Generation of Mutants of Nuclear-Encoded Plastid Proteins Using CRISPR/Cas9 in the Diatom Phaeodactylum tricornutum.- In Silico Tools for the Prediction of Protein Import into Secondary Plastids.- AT_CHLORO - The First Step When Looking for Information about Subplastidial Localization of Proteins.
Primary Endosymbioses: Emergence of the Primary Chloroplast and the Chromatophore, Two Independent Events.- Complex Endosymbioses I: From Primary to Complex Plastids, Multiple Independent Events.- Complex Endosymbiosis II: The Non-Photosynthetic Plastid of Apicomplexa Parasites (The Apicoplast) and Its Integrated Metabolism.- Diversity and Plasticity of Plastids in Land Plants.- The Main Functions of Plastids.- Plastid Transient and Stable Interactions with Other Cell Compartments.- Imaging Plastids in 2D and 3D: Confocal and Electron Microscopy.- Purification of Chloroplasts and Chloroplast Subfractions: Envelope, Thylakoids, and Stroma - From Spinach, Pea, and Arabidopsis thaliana.- Isolation of Inner and Outer Membranes of the Chloroplast Envelope from Spinach and Pea.- Purification of Non-Green Plastids (Proplastids and Amyloplasts) from Angiosperms, and Isolation of Their Envelope Membranes.- In Vitro Protein Import into Isolated Chloroplasts.- Analysis of the MTL Super-Complex at Contact Sites between Mitochondria and Plastids.- Isolation of Plastid-Fractions from the Diatoms Thalassiosira pseudonana and Phaeodactylum tricornutum.- Isolating the Plasmodium falciparum Apicoplast using Magnetic Beads.- Extraction and Quantification of Lipids from Plant or Algae.- Quantitative Assessment of the Chloroplast Lipidome.- Determination of the DNA/RNA-Associated Sub-Proteome from Chloroplasts and Other Plastid Types.- Mapping Plastid Transcript Population by Circular Reverse Transcription-Polymerase Chain Reaction.- Bioinformatic Analysis of Chloroplast Gene Expression and RNA Post-Transcriptional Maturations using RNA Sequencing.- A Guide to the Chloroplast Transcriptome Analysis using RNA-Seq.- A Toolkit for the Characterization of the Photoprotective Capacity of Green Algae.- Rescue of Deletion Mutants to Isolate Plastid Transformants in Higher Plants.- Genetic Analysis of Chloroplast Biogenesis, and Function and Mutant Collections.- Generation of Mutants of Nuclear-Encoded Plastid Proteins Using CRISPR/Cas9 in the Diatom Phaeodactylum tricornutum.- In Silico Tools for the Prediction of Protein Import into Secondary Plastids.- AT_CHLORO - The First Step When Looking for Information about Subplastidial Localization of Proteins.
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