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Combining a Base Deaminase Mutator with Phage-Assisted Evolution

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Phage Engineering and Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2793))

Abstract

Phage-assisted evolution has emerged as a powerful technique for improving a protein’s function by using mutagenesis and selective pressure. However, mutations typically occur throughout the host’s genome and are not limited to the gene-of-interest (GOI): these undesirable genomic mutations can yield host cells that circumvent the system’s selective pressure. Our system targets mutations specifically toward the GOI by combining T7 targeted mutagenesis and phage-assisted evolution. This system improves the structure and function of proteins by accumulating favorable mutations that can change its binding affinity, specificity, and activity.

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Acknowledgement

We are grateful for assistance from Afnan Khan and Montdher Hussain and for support from NSERC and the UTM Office of Research.

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Authors and Affiliations

  1. Department of Chemistry, University of Toronto, Mississauga, ON, Canada

    Maryam Ali, Raneem Akel, Maria J. Botero & Jumi A. Shin

Authors
  1. Maryam Ali
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  2. Raneem Akel
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  3. Maria J. Botero
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  4. Jumi A. Shin
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Corresponding author

Correspondence to Jumi A. Shin .

Editor information

Editors and Affiliations

  1. Cellular Signaling Laboratory International Research, Center for Sensory Biology and Technology of MOST, Key Laboratory of Molecular Biophysics of MOE, College of Life Science and Technology, Huazhong University of Science and Technology (HUST), Wuhan, Hubei, China

    Huan Peng

  2. College of Life Science and Technology, Huazhong University of Science and Technology (HUST), Wuhan, Hubei, China

    Jianfeng Liu

  3. Department of Chemical and Biomolecular Engineering, Department of Chemistry and Biochemistry, University of California, Los Angeles, CA, USA

    Irene A. Chen

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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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Ali, M., Akel, R., Botero, M.J., Shin, J.A. (2024). Combining a Base Deaminase Mutator with Phage-Assisted Evolution. In: Peng, H., Liu, J., Chen, I.A. (eds) Phage Engineering and Analysis. Methods in Molecular Biology, vol 2793. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3798-2_4

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  • DOI: https://doi.org/10.1007/978-1-0716-3798-2_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3797-5

  • Online ISBN: 978-1-0716-3798-2

  • eBook Packages: Springer Protocols

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