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21 thoughts on “S1 – Thron, Tran, Sacconi”
Why is the possibility of mass producing terpenes important?
Thanks for your question, Terpenes are incredibly useful compounds for therapeutics and beauty. Therefore it is important to be able to easily produce large amounts of these compounds in ways that will not wipe out their natural sources, and this is one such way that we can do that!
Which cancer therapeutic is Humulene a precursor?
Thank you for your question, it is the precursor for a therapeutic called Zerumbone!
Why were soybeans chosen as the high yield crop for the production of alpha humulene?
Thank you for your question. Soy beans were selected because there has been a good amount of previous research on their biological pathways and they are easy to grow in large quantities!
What is the potential impact of increasing the alpha humulene?
Very nice project! Would the CRIPS-Cas9 system have worked for means of insertion? Also, what is the super-coiling effect?
Is it clear where the contamination in the last gel could have come from?
Why do you insert the HUM-opt sequence into the B/W section?
Thank you for your question, we implement it into that sequence in order to select for the bacteria that are holding our gene. Basically, if the B/W sequence is intact, it will grow blue colonies, but if it is broken, it will grow white ones. So we can select the white colonies off the plate and they are more likely to contain our gene!
Thanks for your question! We inserted the HUM sequence into the B/W section in order to visualize if the assembly happened as expected. If the B/W section was removed, our plates would show white colonies.
What is the super coiling effect and what causes it?
Thank you for your question, the super-coiling effect occurs with circular DNA from bacterial plasmids. Basically, when the DNA is all intact in the plasmid and it is unbroken, some coiling occurs which condenses the size of the DNA. Therefore when we run it on the gel, it can travel slightly further down and doesn’t reflect the actual length of the plasmid sequence.
The supercoiling effect makes bands look artificially small due to a plasmid being circular.
This makes DNA look artificially small because a plasmid is circular.
Very interesting project! How would you begin the process of integrating this gene into the soy bean plant?
Thanks for your question, the next steps would be to integrate our current segment into an agrobacterium vector which can infect the plants and add our promoter, gene, and terminator to the plant.
After we are able to insert our coding region, promoter and terminator into our desired vector, we can put the final vector into agrobacterium and then transfer this to the soybeans.
Do you have any theories on where those two unaccounted bands in the gel might have arisen from?
We are not really sure. I think it could have been from experimental error during the miniPrep DNA isolation, or a failure of sterilization when plating.