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Transposon Insertion Mutagenesis in Hyperthermophilic Crenarchaeon Sulfolobus islandicus

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2522))

Abstract

Transposon insertion mutagenesis is a forward genetic approach that has been widely utilized for genetic characterization of bacteria and single-celled eukaryotes, and its applications are being rapidly expanded into a few archaeal model organisms for gene function analysis. Previously, we developed a Tn5-based in vivo transposon insertion mutagenesis system in the hyperthermophilic crenarchaeon S. islandicucs M.16.4 and defined the essential gene set under laboratory growth conditions. In this chapter, we will mainly focus on presenting details regarding the generation of a near-saturating transposon insertion mutant library in this crenarchaeal model. We envision that the traditional transposon-based forward mutagenesis screening paired with next generation sequencing will greatly speed up the exploration of archaeal genomic features.

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Acknowledgments

C.Z. and R.J.W are supported by a grant from US National Science Foundation (DEB: 1355171). We thank Dr. Ahmed M. Abdel-Hamid, John R. Schneider, Ruben L. Sanchez-Nieves, Yuan Li, and Rebecca L. Wipfler for carefully reading and editing the manuscript.

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Authors and Affiliations

  1. Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Champaign, IL, USA

    Changyi Zhang & Rachel J. Whitaker

  2. Department of Microbiology, University of Illinois at Urbana-Champaign, Champaign, IL, USA

    Changyi Zhang & Rachel J. Whitaker

Authors
  1. Changyi Zhang
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  2. Rachel J. Whitaker
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Correspondence to Changyi Zhang .

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Editors and Affiliations

  1. Cellular Biochemistry of Microorganisms - Biochemie III, Universität Regensburg, Regensburg, Bayern, Germany

    Sébastien Ferreira-Cerca

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Zhang, C., Whitaker, R.J. (2022). Transposon Insertion Mutagenesis in Hyperthermophilic Crenarchaeon Sulfolobus islandicus. In: Ferreira-Cerca, S. (eds) Archaea. Methods in Molecular Biology, vol 2522. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2445-6_10

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  • DOI: https://doi.org/10.1007/978-1-0716-2445-6_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2444-9

  • Online ISBN: 978-1-0716-2445-6

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