IGEM:MIT/2007/Notebook/2007-6-24

Stuff we found after researching
Semmie: Atheriosclerosis, what it is. Lipase production in E. coli (gene LipA). Probiotics-lactobacillus. Major obstacle: how to make our system work. Other considerations: do we need different facilities to do this project?

Discussion: secretion? how will that work?

how could we test it (in vivo/vitro)?

what are lipid digestion products? (We don't want the lipid to float around)

how do you transfer it from digestive tract to blood?

there are systems that exist to simulate cardivascular environments (velocity, turbulence, blood pressure).

This project sounds cooler, but it has more obstacles that we need more time (probably more time than we have) to handle.

What microorganism would we use this in? our grads are more familiar with e. coli/yeast.

Might not work since we don't have a way to build upon what has been done - we could transfer it over to e. coli but it hasn't been done in e. coli

Toan and Jess: mercury-binding surface display protein and promoter, cadmium and copper resistance genes put into e. coli

Discussion: how cadmium disrupts cells

lead cadmium mercury induced promoters papers, the stuff apparently just diffuses in.

might not need surface display mechanisms

could try just knocking out efflux pumps (will probably kill cell, what other resistance mechanisms exist)

ought to look up reducing enzymes.

Alex: phosphorylation cascades. binding of phosphate group to protein (works like on/off switch) causes others to go on/off. cadA, cadC encoding for cadmium resistance. issues with cadmium pump. reducing ions as a possibility.

Dity: Edinburgh's arsenic project. they basically let arsenic diffuse in, didn't transport it in. looked into cadmium, lead, mercury resistance. arsenate/copper biosensor (inducible promoters). need to find a safe, powerful metal-induced promoter.

Discussion:

bernice found an article about a divalent metal transporter.

Bernice: did a little of everything. Looked into plastic attachment, has a list of peptide sequences we can use. (still looking for another binding just in case). Electric conducting, polypyrolle.

discussion:

bacteria powered micro-rotary motor.

going to use the two strong-binding polystyrene binding peptide sequences

Plan for the week
Monday: present idea to grads construct our orders - need DNA sequences for: display protein insert (minus peptide to be displayed), 2 polystyrene peptide, use computer program to make oligos place orders make primers

Tuesday: primer design brainstorm for ppt on thursday

rest of week: leave open