<p>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.</p>
<p>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.</p>
<p>Read more on our Materials and Methods page</p>
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<divclass="h1">UCYN-A isolation</div>
<p>To facilitate the study of UCYN-A, a protocol to isolate the organelle from B. bigelowii is required. This has been done before, relying on a Percoll gradient. We attempted to develop an alternative protocol for future research via cell sorting which should prove quicker and less laborious, for future teams to be able to use.</p>
<p>Read more on our <ahref="https://2024.igem.wiki/tu-delft/results">Results</a> page </p>
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<divclass="h1"><em>B. bigelowii</em> on list A1</div>
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<p>B. bigelowii is an organism of great interest thanks to UCYN-A. This is because the nitroplast is something like an “evolutionary snapshot” of organellogenesis, and its capability to fix nitrogen makes B. bigelowii the only known nitrogen-fixing eukaryote. However, B. bigelowii is not a model organism, and as a coccolithophore with no biosafety class assigned by any institution, it can prove complicated to gain authorization to work with it. Meanwhile, many experiments on our roadmap for the future of nitroplast transplantation involve modifying B. bigelowii, so this presented a problem.
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<p>We have worked together with our department's biosafety officer to submit a formal request to the Dutch National Institute for Public Health and the Environment (RIVM) for B. bigelowii to be approved into list A1. Organisms in this list are considered equal to common model organisms like E. coli in terms of biosafety and so they can be genetically modified in BSL-1 labs, insofar as no hazardous sequences are involved. We believe this classification will facilitate future research on B. bigelowii by Dutch teams immensely and should make it easier for teams from other countries to get clearance on experiments as well.</p>
