Meeting Minutes for July 7, 2006
From 2006.igem.org
| Line 7: | Line 7: | ||
-MotB flagellar machinery is intact, not structurally changing bacteria, just the motor (good) | -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 | - 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 | '''Sender cell:''' all parts grown up, minipreps on all parts. Colonies didn’t grow | ||
| Line 31: | Line 30: | ||
-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 | -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) | -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''' | '''Magnetotacticbacteria''' | ||
| Line 52: | Line 43: | ||
-will try plating the bacteria now that we have access to anerobic chamber | -will try plating the bacteria now that we have access to anerobic chamber | ||
| - | |||
| - | |||
| - | |||
| - | |||
'''Tasks that members have taken on:''' | '''Tasks that members have taken on:''' | ||
| Line 74: | Line 61: | ||
'''Future Planning: Jason, Brendan, John Cumbers''' | '''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 | ||
Revision as of 19:06, 7 July 2006
Minutes for Weekly meeting (7/7)
Indiv. Member Responsibilities: still underway
Freeze tag: Problems: -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 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 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
