Characterising a new Asgard archaeon form an early-Earth analogue site.
The first chapter of my PhD involved trialing different conditions to refine the cultivation of a notoriously difficult-to-isolate microbe - the Asgard Archaea. This was followed by characterising cytoskeletal and morphological proteins using a new mixed sequence-based and structure-based annotation approach alongside a comparison with the other 3 cultured representatives. The new organism, Nerearchaeum marumarumayae Loki-ASV2, was cultivated from smooth microbial mats from Shark Bay, Western Australia, a World Heritage-listed site that hosts one of the most diverse systems of stromatolites that are considered analogues of life during the Proterozoic (~ 2 billion years ago). The annotation work features in the published Current Biology research article.

Tubulin Superfamily proteins in the Asgard Archaea.
Tubulin Superfamily proteins are fundamental cytoskeletal proteins found in all domains of life and are foundational to understanding the evolution of microbial life during eukaryogenesis. My bioinformatics-heavy chapter scoured the GlobDB dataset to classify the diversity and distribution of Tubulin Superfamily proteins in the Asgard Archaea and provides a basis for understanding the evolutionary trajectories of prokaryotic-like and eukaryotic-like protein homologues. This work is currently unpublished, so please reach out to me if you would like to discuss my findings.

Tubulin protein biochemistry.
I recombinantly co-expressed and purified eukaryotic-like tubulin proteins from N. marumarumayae in E. coli, using my bespoke 'sequence-harmonised' sequence. Despite a heap of optimisation, I never found an ideal polymerisation conditions before Florian Wollweber and Jingwei Xu go there first in 2025. However, I now have a solid background in plasmid construction, protein expression and purification, enzymatic assays, nanoDSF, mass photometry and Western Blotting. I was also introduced to IFM as we tried to visualise the tubulins in vivo.