Abstract
A well-functioning genetic system, which is important for studying gene functions in vivo, requires a transformation method, a vector system and a selection system. Sulfolobus acidocaldarius is a crenarchaeal model organism that grows optimally at 75 °C and a pH of 3. These extreme growth conditions cause some difficulties in developing a genetic system. With continuous efforts, versatile genetic tools have been developed for different species from the order of Sulfolobales. In this chapter, we describe the methods for the available genetic tools in S. acidocaldarius including a (1) transformation method, (2) pop in/pop out strategy to generate markerless deletion mutants and (3) a plasmid-based expression system.
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Acknowledgements
We are grateful to all the bachelor’s, master’s, and PhD students; technicians; and postdoctoral researchers who helped developing and optimizing the genetic system for Sulfolobus acidocaldarius throughout the years, especially Michaela Wagner and Alexander Wagner. MvW received support from the Momentum grant 94933 from the Volkswagen Foundation. XY was funded by the Life? Grant Az 96727 from the Volkswagen Foundation, and AR received support from the HotAcidFactory grant 031B0848C from the BMBF (Federal Ministry of Education and Research).
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Ye, X., Recalde, A., Albers, SV., van Wolferen, M. (2022). Methods for Markerless Gene Deletion and Plasmid-Based Expression in Sulfolobus acidocaldarius. 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_8
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DOI: https://doi.org/10.1007/978-1-0716-2445-6_8
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