Chloroplast Research in Arabidopsis: Methods and Protocols, Volume I

ISBN-13: 9781617792335 / Angielski / Twarda / 2011 / 386 str.

R. Paul Jarvis

Opis

Chloroplasts are essential for the survival and flourishing of life on Earth. Over the years, chloroplast biology has been studied in a variety of different organisms, leading to the significant disadvantage that findings which were made by using different experimental systems or species were not always directly cross-comparable. The relatively recent adoption of Arabidopsis thaliana as the model organism of choice for plant science research, across the globe, has led to its emergence as a pre-eminent system for research on chloroplasts and other types of plastid. In Chloroplast Research in Arabidopsis: Methods and Protocols, expert researchers bring together some of the most important, modern techniques and approaches for chloroplast research, with the unifying theme of Arabidopsis as the model system. Volume I explores topics such as genetics, cytology, in vivo analysis, gene expression, and protein accumulation, as well as protein transport, localization, and topology. 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.Authoritative and convenient, Chloroplast Research in Arabidopsis: Methods and Protocols serves as an ideal reference for all researchers with a general interest in chloroplasts, plastids, or related processes.

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Part I: Genetics, Cytology, and In Vivo Analysis

 

1. Screening or Selection for Chloroplast Biogenesis Mutants of Arabidopsis, Following Chemical or Insertional Mutagenesis

            Enrique López-Juez and Alison Hills

 

2. Analysis of Plastid Number, Size, and Distribution in Arabidopsis Plants by Light and Fluorescence Microscopy

            Kevin Pyke

 

3. Immunofluorescence Microscopy for Localization of Arabidopsis Chloroplast Proteins

            Stanislav Vitha and Katherine W. Osteryoung

 

4. Transient Expression and Analysis of Chloroplast Proteins in Arabidopsis Protoplasts

            Dong Wook Lee and Inhwan Hwang

 

5. Visualisation of Stromules on Arabidopsis Plastids

            John C. Gray, James A. Sullivan, and Christine A. Newell

 

6. Analysis of Chloroplast Movement and Relocation in Arabidopsis

            Masamitsu Wada and Sam-Geun Kong

 

7. Studying Starch Content and Sedimentation of Amyloplast Statoliths In Arabidopsis Roots

            John Stanga, Allison Strohm, and Patrick H. Masson

 

8. Studying Arabidopsis Chloroplast Structural Organization Using Transmission Electron Microscopy

            Stefan Hyman and R. Paul Jarvis

 

9. Transplastomics in Arabidopsis: Progress Towards Developing an Efficient Method

            Kerry Ann Lutz, Arun Azhagiri, and Pal Maliga

 

Part II: Gene Expression and Protein Accumulation

 

10. Isolation, Quantification, and Analysis of Chloroplast DNA

            Beth A. Rowan and Arnold J. Bendich

 

11. Measurement of Transcription Rates in Arabidopsis Chloroplasts

            Yan O. Zubo, Thomas Börner, and Karsten Liere

 

12. Studying the Structure and Processing of Chloroplast Transcripts

            Alice Barkan

 

13. In Vitro RNA Binding Assay for Studying Trans-Factors for RNA Editing in Chloroplasts

            Toshiharu Shikanai and Kenji Okuda

 

14. Studying Translation in Arabidopsis Chloroplasts

            Paolo Pesaresi

 

15. Studying Proteases and Protein Turnover in Arabidopsis Chloroplasts

            Lars L.E. Sjögren and Adrian K. Clarke

 

Part III: Protein Transport, Localization, and Topology

 

16. In Silico Methods for the Identification of Organellar and Suborganellar Targeting Peptides in Arabidopsis Chloroplast Proteins and for Predicting the Topology of Membrane Proteins

            Sandra Tanz and Ian Small

 

17. Rapid Isolation of Arabidopsis Chloroplasts and Their Use for In Vitro Protein Import Assays

            Henrik Aronsson and R. Paul Jarvis

 

18. Energetic Manipulation of Chloroplast Protein Import and the Use of Chemical Cross-Linkers to Map Protein-Protein Interactions

            Hitoshi Inoue, Fei Wang, Takehito Inaba, and Danny J. Schnell

 

19. Isolation of Arabidopsis Thylakoid Membranes and Their Use for In Vitro Protein Insertion or Transport Assays

            Thomas Bals and Danja Schünemann

 

20. Determining the Location of an Arabidopsis Chloroplast Protein Using In Vitro Import Followed by Fractionation and Alkaline Extraction

            Chiung-Chih Chu and Hsou-min Li

 

21. Studying Arabidopsis Envelope Protein Localization and Topology Using Thermolysin and Trypsin Proteases

            John Froehlich