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Phage Display pp 519–534Cite as

Metasecretome Phage Display

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1701))

Abstract

Metasecretome is a collection of cell-surface and secreted proteins that mediate interactions between microbial communities and their environment. These include adhesins, enzymes, surface structures such as pili or flagella, vaccine targets or proteins responsible for immune evasion. Traditional approaches to exploring matasecretome of complex microbial communities via cultivation of microorganisms and screening of individual strains fail to sample extraordinary diversity in these communities, since only a limited fraction of microorganisms are represented by cultures. Advances in culture-independent sequence analysis methods, collectively referred to as metagenomics, offer an alternative approach that enables the direct analysis of collective microbial genomes (metagenome) recovered from environmental samples. This protocol describes a method, metasecretome phage display, which selectively displays the metasecretome portion of the metagenome. The metasecretome library can then be used for two purposes: (1) to sequence the entire metasecretome (using PacBio technology); (2) to identify metasecretome proteins that have a specific function of interest by affinity-screening (bio-panning) using a variety of methods described in other chapters of this volume.

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Acknowledgment

This work was funded by the New Zealand Ministry of Business, Innovation and Employment (contract C10X0803), The Royal Society of New Zealand through a Marsden Fast Start grant and Palmerston North Medical Research Foundation. M.C. was partially supported by the Institute of Fundamental Sciences (Massey University, New Zealand) and F.N. was supported by a Commonwealth Fellowship and a Massey University Bursary. We would like to thank DSMZ Sequencing facility (Germany) and Dr. Boyke Bunk for help in development and optimization of protocol for PacBio sequencing of phage-display libraries.

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Authors and Affiliations

  1. Institute of Fundamental Sciences, Massey University, Private Bag 11 222, Palmerston North, 4442, New Zealand

    Milica Ciric, Filomena Ng, Jasna Rakonjac & Dragana Gagic

  2. Grasslands Research Centre, AgResearch Ltd., Palmerston North, New Zealand

    Filomena Ng

Authors
  1. Milica Ciric
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  2. Filomena Ng
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  3. Jasna Rakonjac
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  4. Dragana Gagic
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Corresponding author

Correspondence to Dragana Gagic .

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Editors and Affiliations

  1. Technische Universität Braunschweig, Braunschweig, Germany

    Michael Hust

  2. Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Minden, Penang, Malaysia

    Theam Soon Lim

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Ciric, M., Ng, F., Rakonjac, J., Gagic, D. (2018). Metasecretome Phage Display. In: Hust, M., Lim, T. (eds) Phage Display. Methods in Molecular Biology, vol 1701. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7447-4_29

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  • DOI: https://doi.org/10.1007/978-1-4939-7447-4_29

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7446-7

  • Online ISBN: 978-1-4939-7447-4

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