Difference between revisions of "Team:Aix-Marseille/Results"

On the right, we tested the protein production using a SDS page and comassie in a Escherichia coli Tg1 strain after transformation with Bba_K1951011. On the right column, it has been observed the protein production without induction with 0.02% arabinose. On the left column, protein production was tested after induction. Results showed the production of the 4 proteins DesA, DesB, DesC and DesD, all involved in the desferrioxamine biosynthesis pathway. On the left, we had a look of the cadaverine production by the lysine decarboxylase DesA. Production has been detected by HPLC using C18 column after induction of the strain. Escherichia coli Tg1 strain was used as a wild type. Cadaverine production has been detected in this strain. In a Tg1 cadA mutant, cadaverine was also produced in a least quantity showing that an other pathway is responsible for the production of cadaverine. In the cadA mutant complemented by Bba_K1951004, the amount of cadaverine was recovered and even beyond the wild type production. Moreover, in the cadA mutant complemented by Bba_K1951011, the cadaverine level produced was even over the wild type and complemented Bba_K1951004 production

We investigated if swimming motility was recovered by a knockout FliC strain. To test complementation with our biobrick strains were stabbed into soft (Claire check 0.5%) LB agar plates, and incubated at 37°C for 4 hours. Three strains are shown: Escherichia coli W3110 wild-type strain, which has a good swimming capacity (lower right); a fliC deletion mutant of W3110 (lower left); and the fliC mutant complemented with BBa_K1951008(top).

Maquette of our process in the futur

Igem Aix Marseille 2016 in a sustainable circle

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 ===Proof of protein production===
-[[File:T--Aix-Marseille--comassieflic.jpeg|left|400px|thumb|Test of our biobrick proteins production using a SDS page and comassie]]
-We investigated if the FliC protein was well produced by our biobrick using SDS PAGE.
-To to do this we performed SDS PAGE and stained with coomassie blue using cells containing this biobrick in plasmid backbone SDS page and coomassie blue.
-From an over night starter, cells were diluted and grown from Abs(600nm)=0.2 to Abs(600nm)=0.6.
-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.
-SDS page and coomassie blue The mixture was loaded onto a polyacrylamide gel and migrated during 50min at 180V.
-Staining was done using coomassie blue.
-The FliC is at mass 51,3kDa, and can be clearly seen in the gel photograph.
 === Proof of fonctionnality===

Difference between revisions of "Team:Aix-Marseille/Results"

Revision as of 21:17, 18 October 2016

Contents

Result

Mobilisation result

Proof of protein production

Proof of fonctionnality

In the futur

Biosorption result

Proof of swimming recovery

Microscopie of the flagellum

In the futur

Improvement of FliC E. coli [http://parts.igem.org/Part:BBa_K1463604 BBa_K1463604]

Project achievement in a table

Futur plan for our project

Project Achievements