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Contribution
Introduction: NitroBLAST as a foundation for novel nitrogen fixation research

Biological nitrogen fixation has served as an inspiration for iGEM teams to build their projects around for many years, and as long as no solution is found to the nitrogen crisis, we believe many more will contribute towards solving it. We conceived NitroBLAST as a foundational project: a way to start a line of research on this exciting and new possibility to engineer a biological nitrogen fixation system. There is so much left to do on the road to creating nitrogen-fixing eukaryote strains, and in the spirit of iGEM, we documented all we have learned from our research as well as interviewing numerous experts, to provide tools and guidance for future teams to pick up where we have left off.

B. bigelowii and UCYN-A omics analysis

UCYN-A is actively undergoing genome reduction as part of its evolution towards an organelle: the reason it cannot live independently is that essential proteins for its survival are no longer present in its genome, but are now encoded in B. bigelowii’s and then imported into UCYN-A. Using recently released proteomics data on B. bigelowii and UCYN-A, along with older genomics and transcriptomics data, we have identified a putative list of proteins that are imported into the organelle. This data provides a solid foundation for further research into which proteins are essential, as we suspect many are redundant. Identifying a list of essential host-encoded proteins is crucial to successfully transplanting UCYN-A into a new host.

We have also created a new transcriptome assembly of B. bigelowii based on raw data from previous studies, using improved algorithms. This allowed us to create a new predicted proteome. We are making all of our omics data available for future iGEM teams along with documentation.

Read more on our Results page

UCYN-A transit peptide

UCYN-A isolation

Peptide binding model

Based on the previously identified transit peptide and the predicted B. bigelowii proteome, we attempted to find any proteins which interact with uTP and thus might play a role in the UCYN-A protein import system. To achieve this, we began developing a proteome-scale peptide-binding prediction tool, which, to our knowledge, does not yet exist.

Unfortunately, due to the strict time constraints in iGEM, we were not able to finish work on this tool, but we are making our existing code and data available to aid any future teams.

Read more on our Materials and Methods page

B. bigelowii on list A1

References
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