Difference between revisions of "Team:Aix-Marseille/Composite Part"

(BBa_K1951011 : Desferrioxamine B producer pathway Streptomyces coelicolor producer)
(BBa_K1951011 : Desferrioxamine B producer pathway Streptomyces coelicolor producer)
Line 52: Line 52:
  
 
* Desferrioxamine B may modulate expression and release of inflammatory mediators by specific cell types.
 
* Desferrioxamine B may modulate expression and release of inflammatory mediators by specific cell types.
 +
 +
==== Results ====
 +
 +
[[File:T--Aix-Marseille--result4.jpeg|500px|right|thumb|Test of our biobrick proteins production using a SDS page and comassie. - : no induction ; + : induction with 0.02% arabinose at Abs(600nm)=0.4]]
 +
We investigated if the DesA, DesB, DesC and DesD proteins were well produced by our biobrick using SDS PAGE.
 +
 +
To do this we performed SDS PAGE and stained with comassie blue using cells containing this biobrick in plasmid backbone [https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols Find here the protocol] From an over night starter, cells were diluted and grown from Abs(600nm)=0.2 to Abs(600nm)=1.
 +
Then 1UOD of cells (1.67ml at 0.6OD) was collected and centrifuged at 5000g for 5min.
 +
After removal of the supernatant, the cell pellet was resuspended in 50µL SDS-PAGE sample buffer.
 +
We heated the mix at 95°C during 15min
 +
[https://2016.igem.org/Team:Aix-Marseille/Experiments/Protocols Find here the protocol]
 +
The mixture was loaded onto a polyacrylamide gel and migrated during 50min at 180V.
 +
Staining was done using coomassie blue.
 +
 +
We compared the production of proteins in different background :
 +
 +
- <i>E. coli</i>  Tg1 strain without any plasmide (negative temoin)
 +
 +
- <i>E. coli</i>  Tg1 strain with pSB1C3 containing the RFP coding sequence (negative temoin)
 +
 +
- <i>E. coli</i>  Tg1 strain complemented with our biobrick Bba_K1951011 before and after induction. (you can observe the production of the 4 proteins on the figure below on the left; right : before induction, left : after induction)
 +
 +
Results showed the production of the 4 proteins DesA, DesB, DesC and DesD, all involved in the desferrioxamine B biosynthesis pathway.
  
 
==Biosorption parts ==
 
==Biosorption parts ==

Revision as of 17:44, 19 October 2016