Abstract
Haloarchaea have evolved to thrive in hypersaline environments. Haloferax volcanii is of particular interest due to its genetic tractability; however, few in vivo reporters exist for halophiles. Haloarchaeal proteins evolved characteristics that promote proper folding and function at high salt concentrations, but many mesophilic reporter proteins lack these characteristics. Mesophilic proteins that acquire salt-stabilizing mutations, however, can lead to proper function in haloarchaea. Using laboratory-directed evolution, we developed and demonstrated an in vivo luciferase that functions in the hypersaline cytosol of H. volcanii.
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Abbreviations
- CDS:
-
Coding sequence
- CBG:
-
Click Beetle Green luciferase
- CCD:
-
Charge-coupled Device
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Acknowledgements
This research was initiated with funding from the NIH (R37GM067152, R01GM107434, and R21MH116150 to CHJ) and completed with funding from MTSU Center for Molecular Biosciences (to JBR). We thank Thorsten Allers (U. of Nottingham) for providing us with strain H1209 and plasmid pTA962, as well as culturing/transformation advice.
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Davis, C.R., Johnson, C.H. & Robertson, J.B. A bioluminescent reporter for the halophilic archaeon Haloferax volcanii. Extremophiles 24, 773–785 (2020). https://doi.org/10.1007/s00792-020-01193-x
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DOI: https://doi.org/10.1007/s00792-020-01193-x