Transposon Insertion Mutagenesis in Hyperthermophilic Crenarchaeon Sulfolobus islandicus

Changyi Zhang, Rachel J. Whitaker

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages163-176
Number of pages14
DOIs
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2522
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Archaea
  • Sulfolobus islandicus
  • Tn5 transposase
  • Transposon insertion library
  • random mutagenesis

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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