Methods for Markerless Gene Deletion and Plasmid-Based Expression in Sulfolobus acidocaldarius

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.

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

Link to chapter: https://link.springer.com/protocol/10.1007/978-1-0716-2445-6_8