Meeting Minutes for July 7, 2006

From 2006.igem.org

Minutes for Weekly meeting (7/7)

Indiv. Member Responsibilities: still underway

Freeze tag:
-MotB flagellar machinery is intact, not structurally changing bacteria, just the motor (good)
- it bridges into to the cell, shouldn’t cause a quantum increase

Sender cell: all parts grown up, minipreps on all parts. Colonies didn’t grow run another gel, digest it, get it sequenced and ligate the 3 parts together.
-will sequence after first digestion (Brendan)
-designing the primers: Annie and Prof. Wessel
Receiver cell: need to run digest of the two parts (Victoria and Megan)
“Freeze Machine”: need to incubate cells, do minipreps (Jamie and Annie)
Unfreeze: do miniprep tonight (Jason and Azeem)

Modeling Mechanism
-put a gene then a protein with a line connecting it, RBS and primer is an effector of that pathway

Problems: can’t inhibit the rate the promoter outputs
-talks about having a metabox
-website: bio tapestry

What are hopes of model?
-10 internal signaling pathways, worth running time courses seeing how stable system is
-does system break down if we vary concentrations?
-is there a way to do positive control, repressalator
-To check that logical sensible
-want to see if signaling cell will actually stop the receiver cell, if not, can we figure out the strength of the RBS necessary to get the job done
-focus of small steps: once we get the sensor and sender of AHL working we can then test it out to define rate laws (do so by putting GFP at end of sequences-to test the POPs)

Magnetotacticbacteria

-still alive
-looked at most concentrated vile, 3 cells in each visual field 200microliters, 50 microliters of food
-divided to some extent
-mixed up another vile to grow in anerobic bacterial incubator
-Hyato requested 4 types of plasmid, that Luciferase expresses much better, email has been sent out -Plamids modified in D5Halpha cells
-can electroborate into bacterial cells and the other to conjugate them
- before we can do anything we need to get a stable stock of the bacteria
-will try plating the bacteria now that we have access to anerobic chamber

Tasks that members have taken on:

-Megan will look at options to synthesize avadin from scratch. Costs ? Practicality ?
-end goal to make a standard shuttle vector that can be used.

-take origin of replication from plasmids that we are looking for and insert them into the shuttle vector as a contribution to the biobricks registry. Important to make it compatible with most bacteria used in the project (Jason)

-wiki has been updated by Annie and Jamie NB: you can update the particular parts for “the Freeze Tag”

General overview of Project (Azeem)

Outreach Coordinator: Megan

Industry Liaison: Victoria

Future Planning: Jason, Brendan, John Cumbers

Questions, Thoughts, Concerns?

(1.)can a biobick could be created for the plasmids?

-we should get a hold of avadin part, then optimize codon usage, find restriction sites
-look at article with Mag A

(2.)look into buying AHL

(3.)How do we test AHL concentration (it is a lactose analog-a simple carbohydrate)?
-Right now we are using TetR promoter, will that be strong enough? Not that well characterisized.
-Brendan suggesting making a biobrick with T4 promoter (wouldn’t be that hard)
-Think we need 0.2 nanomoles of AHL to activate the AHL receiver

(4.)LVA: is it made individually? Cell recognizes it as mRNA