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SMC Complexes

Methods and Protocols

  • Book
  • © 2019

Overview

Editors:
  1. Anjana Badrinarayanan

    1. National Centre for Biological Sciences, Tata Institute of Fundamental Research (TIFR), Bangalore, India

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  • Includes cutting-edge techniques
  • Provides step-by-step detail essential for reproducible results
  • Contains key implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 2004)

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About this book

This book brings together recent methods and theoretical approaches developed to dissect the activity and function of bacterial and eukaryotic Structural Maintenance of Chromosome (SMC) proteins. The protocols explore numerous subjects such as depletion systems to assess SMC function, genetic manipulation of SMC proteins, chromosomal, biochemical, and microscopy-based assays of SMC activity, as well as theoretical modeling of SMC activity. Written for the highly successful Methods in Molecular Biology series, 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 practical, SMC Complexes: Methods and Protocols will greatly aid researchers looking into the mechanism of action of SMC complexes on DNA.


Chapters 10 and 19 areavailable Open Access under a CC BY 4.0 license via link.springer.com.

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Keywords

Table of contents (22 protocols)

  1. Front Matter

    Pages i-xiii
    Download chapter PDF

  2. Depletion Systems to Assess SMC Function

    1. Front Matter

      Pages 1-1
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    2. Using Cell Cycle-Restricted Alleles to Study the Chromatin Dynamics and Functions of the Structural Maintenance of Chromosomes (SMC) Complexes In Vivo

      • Demis Menolfi, Dana Branzei
      Pages 3-16

    3. Degradation of S. cerevisiae Cohesin with the Auxin-Inducible Degron System

      • Clémentine Brocas, Cécile Ducrot, Karine Dubrana
      Pages 17-24

    4. Efficient Depletion of Fission Yeast Condensin by Combined Transcriptional Repression and Auxin-Induced Degradation

      • Yasutaka Kakui, Frank Uhlmann
      Pages 25-33Open Access

    5. Conditional Mutation of SMC5 in Mouse Embryonic Fibroblasts

      • Himaja Gaddipati, Marina V. Pryzhkova, Philip W. Jordan
      Pages 35-46

  3. Genetic Manipulation of SMC Function

    1. Front Matter

      Pages 47-47
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    2. High-Throughput Allelic Replacement Screening in Bacillus subtilis

      • Marie-Laure Diebold-Durand, Frank Bürmann, Stephan Gruber
      Pages 49-61

    3. Identifying Functional Domains in Subunits of Structural Maintenance of Chromosomes (SMC) Complexes by Transposon Mutagenesis Screen in Yeast

      • Avi Matityahu, Michal Shwartz, Itay Onn
      Pages 63-78

    4. Multicomponent Yeast Two-Hybrid System: Applications to Study Protein–Protein Interactions in SMC Complexes

      • Jan Josef Paleček, Lucie Vondrová, Kateřina Zábrady, Jakub Otočka
      Pages 79-90

    5. Knocking in Multifunctional Gene Tags into SMC Complex Subunits Using Gene Editing

      • Paul Kalitsis, Tao Zhang, Ji Hun Kim, Christian F. Nielsen, Kathryn M. Marshall, Damien F. Hudson
      Pages 91-102

  5. Biochemical Assays of SMC Activity

    1. Front Matter

      Pages 167-167
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    2. Dissecting DNA Compaction by the Bacterial Condensin MukB

      • Rupesh Kumar, Soon Bahng, Kenneth J. Marians
      Pages 169-180

    3. In Vivo and In Vitro Assay for Monitoring the Topological Loading of Bacterial Condensins on DNA

      • Koichi Yano, Koichiro Akiyama, Hironori Niki
      Pages 181-196

    4. A Protocol for Assaying the ATPase Activity of Recombinant Cohesin Holocomplexes

      • Menelaos Voulgaris, Thomas G. Gligoris
      Pages 197-208
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Editors and Affiliations

  • National Centre for Biological Sciences, Tata Institute of Fundamental Research (TIFR), Bangalore, India

    Anjana Badrinarayanan

Bibliographic Information

  • Book Title: SMC Complexes

  • Book Subtitle: Methods and Protocols

  • Editors: Anjana Badrinarayanan

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-4939-9520-2

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC, part of Springer Nature 2019

  • Hardcover ISBN: 978-1-4939-9519-6Published: 31 May 2019

  • Softcover ISBN: 978-1-4939-9522-6Published: 14 August 2020

  • eBook ISBN: 978-1-4939-9520-2Published: 30 May 2019

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XIII, 336

  • Number of Illustrations: 36 b/w illustrations, 55 illustrations in colour

  • Topics: Genetics and Genomics, Protein Science

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