Difference between revisions of "Team:Toronto/Notebook-w02-fri-2"

 
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Latest revision as of 06:57, 19 October 2016

console.js:26

Week 2: May 27, 2016

Friday, 5/27
LynnMembers present: Celine, Tam, Hamed, Cathy, DK, Alex, Katariina
Lab Stuff: (research, plasmid design, protocol design, troubleshooting)
Research into the appropriate promoter:
"Transcription of gesABC is delayed compared with the Au-induced expression of golTS or golB, and requires higher GolS intracellular levels, suggesting a hierarchical activation of the gol regulon to cope with contaminated environments."
In the presence of gold ions: There is a sensitivity/specificity trade-off between using pgolB and pges (http://www.ncbi.nlm.nih.gov/pubmed/17919284: see below)
A
B
C
1
pgolBpges
2
Sensitivity (i.e. Transcription level in presence of gold(III) ions)Higher (36 folds) i.e. easier to set offLower i.e. harder to set off
3
Specificity (i.e. Response to Cu(II))Lower i.e. more sensitive to Cu(II)Higher (65 folds) i.e. less sensitive to Cu(II)
Table1
Sensitivity:
clipboard_2016-05-27_11:31:47.png
thumbnail
Time-course of GolS induction of gesABC and golB transcription in the presence of Au(III) (AuHCl4). The maximum amount of transcription response shows 278 Miller units for gesABC and 10090 Miller units for golB - 36-fold difference. There is an indication that pGesABC was less sensitive to pGolB in the presence of Au(III) as pGesABC transcription level was significantly low in the first 5 hours. Transcription reaches the maximum capacity at 5.8 hours for gesABC and 3.6 hours for GolB.
Specificity:
clipboard_2016-05-27_11:15:44.png
thumbnail
GesABC::LacZ fused gene expresses no response to the addition of CuSO4 (Cu(II)) in the system. GolB::LacZ fused gene expresses a 65-fold response to the addition of CuSO4 (Cu(II)) in the system.
In general, there is a high level of sensitivity to Au ions in pGolB than in pGesABC. However, pGesABC shows a higher specificity to Au ions than Cu ions in comparision to pGolB which can be induced by both Au and Cu ions.
Creation of golS variants in the feature library using the following table to assist in optimizing the new codons:
E. coli Codon Bias.gif
thumbnail
Administrative: (shipment orders, inventory)
Email Updates: (Correspondance with proffessors, shipping companies, etc)
Followed up regarding lockers
Drafted e-mail to Kayla and sent to Umar for review before sending it out
Tasks to Follow up on - leading into week 3
Promoter debacle (specificity/sensitivity) - All
Luciferase/RFP (reaction with cellulose) - Kat
merR family and Nickle sensors generally (as back up) - Wiki team
Outlining Dr. Pardee's updated protocol - Kat
golS structures (4 versions) - Tam
118 - Kat
113 - Alex
113 + 118 - Alex
CupR - Cathy
Tasks leading into week 3 - v2.jpg
thumbnail
Meeting Notes:
Weekly meetings in WB 407, hopefully starting from the week that we are in the lab, this will be on Mondays at ? time
We need a common medium of communication. – We decided on Facebook.
We got the credit cards, which was given to DK/Zarifah. If you need to spend anything talk to Adele.
PnP have yet to finish the synenergene project. They will now focus on heavy metal biosensing (as a environmental and health and safety concern), and bringing up wet lab and GolS as a proof-of-concept example. Furthermore, they will have Zika virus on the side as an example of an application for paper based biosensing methods.