Team:Northwestern/Notebook/WeekTwo
From 2012.igem.org
Week Two - 24 June to 30 June
Monday
Researched more project ideas: phytoremediation of heavy metals and other pollutants in the soil, bacterial signaling chain, bioavailability of iron in food
Bacterial signaling chain idea is extremely well developed in a powerpoint
Advisor meeting:
Discussed bacterial signaling chain idea in detail
The professors had reservations about the novelty of the project as some teams have done similar light communication ideas
The professors suggested looking up flash vs glow luciferase kinetics for more information regarding optimal activation of the bacteriorhodopsin/halorhodopsin
Methanotrophs:
the low expression of the gene in vivo makes the project difficult
look into MAGE codon optimization
Fire alarm cut meeting short, so another meeting is planned tomorrow
Tuesday
Continued research regarding phytoremediation of heavy metals, bacterial signaling chain, and bioavailability of nutrients in food
Jen and Advisor Meeting:
Completing the methanotroph project would require using a lot of MAGE and is not very suitable for an iGEM project. The project is also very risky as it might just not work and we would have very little to show for it.
Phytoremediation is troubling the advisors: they both do not like using plants and are not sure about how useful phytoremediation is as it is never utilized in practice due to a very slow remediation rate.
The light communication signaling chain project is well developed and is a definite possibility. The professors are slightly worried if one bacteria lighting up will be strong enough to trigger the next bacteria.
The bioavailability of iron project however is going strong. The advisors liked how the system was well characterized but also novel to igem .
They suggested a probiotic aspect to the project as a delivery system.
The advisors suggested to research one project in extreme detail for the next 24 hours.
We choose the bioavailability of iron and other nutrients to research more. Specifically, developing the probiotic and secretion ideas more fully.
Wednesday
Continued research regarding the project idea, mainly the bioavailability of iron.
Advisor meeting:
We decided on our project: Phytase production in e. coli to make iron bioavailable!
Research is still needed to finalize the list of parts needed to complete the project.
Thursday
Research for the phytase probiotic production project was continued. Two major problems need to be solved with the project:
1. A delivery or secretion system to export the phytase enzyme out of the e. coli and into the gut.
(continue research regarding stochastically inducible promoters and bistable switches)
2. If iron absorption can occur outside of the duodenum, where little to no e. coli live
Tae and Lajja made LB plates(mixing agar and media)
Grant worked on stochastic promoters and methods of ingestion while Mike worked on the mechanism of phytate breakdown
Lajja continued asking for supplies from our sponsors. Dr. Mordacq had a meeting with Lajja about supplies needed that are critical.
Yuan cleaned out the refrigerator
Friday
Continued researching parts for the project.
Brian transformed DNA(RFP) into our competent cells
Brian streaked noncompetent cells on a plate and put them in the cold room, preparing to make new competent cells for next week.
Saturday
Brian counted the individual transformed colonies to check to see if the first set of cells are competent
We do not think the cells are as competent as needed, 10^6 / 10^7 colonies per plate is ideal. The plates made had approximately 2.5^5 colonies per plate.
New super competent cells are needed as these cells were not competent enough