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Protein–Protein Interactions: Bimolecular Fluorescence Complementation and Cytology Two Hybrid

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Bacterial Secretion Systems

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2715))

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Abstract

Identifying protein–protein interactions between machine components of bacterial secretion systems and their cognate substrates is central to delineating how the machines operate to translocate their substrates. Further, establishing which among the machine components and their substrates interact with each other facilitates (i) advancement in our understanding of the architecture and assembly of the machines, (ii) understanding the substrates’ translocation routes and mechanisms, and (iii) how the machines and the substrates talk to each other. Currently, though diverse biochemical methods exist in identifying direct and indirect protein–protein interactions, they primarily remain in vitro and can be quite labor intensive. They also may capture/exhibit false-positive interactions because of barrier breakdowns as part of methodology. Thus, adopting novel genetic approaches to help visualize the same in vivo can yield quick, advantageous, reliable, and informative protein–protein interactions data. Here, we describe the easily adoptable bimolecular fluorescence complementation and cytology-based two-hybrid assays to understand the bacterial secretions systems.

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Acknowledgments

I thank Peter (Prof. Peter J Christie) and Bill (Prof. William Margolin), both from Department of Microbiology and Molecular Genetics, University of Texas Health Science Center, Houston, Texas, USA. Peter was instrumental in mentoring and providing excellent opportunity to train as a postdoc in his lab and Bill for all the help, guidance, and training in fluorescent microscopy.

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Author notes

  1. Authors Dyuti Purkait and Mohd Ilyas have equally contributed to this chapter.

Authors and Affiliations

  1. Translational Health Science and Technology Institute, NCR Biotech Science Cluster, Faridabad, Haryana, India

    Dyuti Purkait, Mohd Ilyas & Krishnamohan Atmakuri

Authors
  1. Dyuti Purkait
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  2. Mohd Ilyas
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  3. Krishnamohan Atmakuri
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Corresponding author

Correspondence to Krishnamohan Atmakuri .

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Editors and Affiliations

  1. Laboratoire d’Ingénierie des Systèmes Macromoléculaires, Institut de Microbiologie de la Méditerranée, Aix-Marseille Univ, CNRS, Marseille, France

    Laure Journet

  2. Laboratoire d’Ingénierie des Systèmes Macromoléculaires, Institut de Microbiologie de la Méditerranée, Aix-Marseille Univ, CNRS, Marseille, France

    Eric Cascales

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Purkait, D., Ilyas, M., Atmakuri, K. (2024). Protein–Protein Interactions: Bimolecular Fluorescence Complementation and Cytology Two Hybrid. In: Journet, L., Cascales, E. (eds) Bacterial Secretion Systems . Methods in Molecular Biology, vol 2715. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3445-5_16

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  • DOI: https://doi.org/10.1007/978-1-0716-3445-5_16

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3444-8

  • Online ISBN: 978-1-0716-3445-5

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