Contribution

Plan for this

ADD THIS LAST, since people need to get the parts into the registry and those numbers need to go on here as well as we can only keep the parts of this that we use for our actual contributions.

Numbers of Parts:
BBa_K5410027

Written portion:

A massive goal of our team is to leave a legacy from the work that we do now. While we are an undergraduate team and we are only able to be in the lab working for under 6 months, we want to create something that can be picked up by another team of researchers that can go on and change the world for the better. Due to this, we have chosen to continue a project from a previous year that we believe can accomplish just that. Our contribution to a future iGEM team is the framework for a project that would give the world a biodegradable plastic made from an interaction between E. coli and algae. This would not just be a contribution to iGEM, but to the world.

In this contribution from our team, not only are we hoping that our progress is picked up by another team that wants to take things further, but we also are providing the international conglomerate of teams with a part and some other prospective parts that we designed. Our currently submitted and most prized part is our kill switch which has the part number PUT KILLSWITCH NUMBER HEREThis kill switch is a composite plasmid part built for E. coli that uses one gene on the plasmid to encode a toxin that would kill the E. coli and another operon-based gene on the same plasmid to encode an antitoxin if a specific molecule is present in the culture. Our team is currently seeking the “Best New Composite Part” award for the creation of this part. The team has also submitted some parts that, while not proven in the lab at this time, they have enough of a design to where the submission team decided to submit it for this year. We will have results for these parts either at the Jamboree or next year.

These parts are as such:

Due to the lack of time we have been able to be in the lab, we are having to prove our other parts work in October. Therefore, we were not comfortable at this time to submit them to the registry in this condition as it is in our belief that we would like to work on them more and possibly submit them next year so that the rest of iGEM can use them when we have a better product. However, we do feel comfortable discussing some of the parts we are creating that could be available in the registry next year from the work we will be able to do within the next few weeks in the lab.

The other currently unfinished parts our team have created the blueprints for both this year and last year, as well as tested to some extent predominantly this year, are: our C. reinhardtii BFP control transformation plasmid, the three C. reinhardtii plasmids coding for the phaA, phaB and phaC enzymes, our acetate pathway knockout plasmids for E. coli, and our ethanol team plasmids. In this contributions section, we shall only cover the ones we believe we will have decent data on by the date of the Jamboree, therefore not all of them will be described in our contributions and we will only showcase the reasons as for why we are building these plasmid parts.

Regarding the plasmid for the control transformation for the algae strain C. reinhardtii, this was a plasmid with a backbone that we found could be used the strain of C. reinhardtii called pChlamy[1] and in that backbone, as of right now, in October we plan on restricting it to then ligate BFP into it to create our recombinant plasmid. Afterwards we shall transform it into C. reinhardtii via electroporation and ensure that it makes the organism glow, confirming our transformation. Our iGEM team hopes that this will eventually end up in the biobrick construct palates sent out each year and will be available to those who are doing genetic research with C. reinhardtii so that they can have a control transformation as soon as they can have it grown. This would, not only cut down on time, but save any team using this a lot of work and hardship that we had to go through to find what plasmid would work with our strain of algae.

Continuing onto some of the other parts that are in production and would likely be of use for future teams, we have the ethanol production plasmid parts and the killswitch plasmid parts. The ethanol production parts are being created to give E. coli the ability to produce ethanol with the intent of causing a stress reaction in the algae inside of our co-culture by that ethanol in order to cause it to produce more of the PHB lipid. Next, the acetate knockout plasmid parts have the purpose of knocking out the ability for E. coli to metabolically synthesize acetate, causing its metabolic pathway to stop at acetyl-CoA. This would cause the E. coli to over produce acetyl-CoA -a metabolite for algae to produce PHB- which would make the cell expel the product to then be taken up by algae in the co-culture and be synthesized by said algae into PHB.

Regardless of the fact that we do not have a finished product in these specific parts, we will probably have better products when the Jamboree comes around and will therefore be able to submit most, if not all, of our constructs to the registry when it reopens in next year’s competition. Even though our current progress would not yield an immediate contribution, in the long term we plan to have quite a few contributions to the parts registry once the kinks are worked out in the laboratory.

While we do not have a lot of completed biobricks and plasmid parts for the registry as of the date of the wiki freeze, our team shall try and get many of them up and working to deliver when we can in the future. Even though our contribution seems small at the time of this freeze, we will have more that can help revolutionize the research of many of the other teams once we have the time to progress our research.

References

[1] Thermofisher. (n.d.). GeneArtTM Chlamydomonas Protein Expression Vector. www.thermofisher.com. https://www.thermofisher.com/order/catalog/product/A24231


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