Difference between revisions of "Team:Aix-Marseille/Results"
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<li><a href="http://polytech.univ-amu.fr/"><img src="http://i.imgur.com/iLFZ36R.png" class="img-grey mautomargin" width="200" height="100"></a></li> | <li><a href="http://polytech.univ-amu.fr/"><img src="http://i.imgur.com/iLFZ36R.png" class="img-grey mautomargin" width="200" height="100"></a></li> | ||
| − | <li><a href="http://www.crous-aix-marseille.fr/social/fsdie-social"><img src="http://i.imgur.com/KlP5G8h.png" class="img-grey mautomargin" width="200" height=" | + | <li><a href="http://www.crous-aix-marseille.fr/social/fsdie-social"><img src="http://i.imgur.com/KlP5G8h.png" class="img-grey mautomargin" width="200" height="150"></a></li> |
<li><a href="http://www.univ-amu.fr/"><img src="http://i.imgur.com/EcfF9Kw.png" class="img-grey mautomargin" width="200" height="100"></a></li> | <li><a href="http://www.univ-amu.fr/"><img src="http://i.imgur.com/EcfF9Kw.png" class="img-grey mautomargin" width="200" height="100"></a></li> | ||
| − | <li><a href="http://www.cnrs.fr/"><img src="http://i.imgur.com/IaWMSIr.jpg" width="200" height=" | + | <li><a href="http://www.cnrs.fr/"><img src="http://i.imgur.com/IaWMSIr.jpg" width="200" height="150" class="img-grey mautomargin"></a></li> |
<li><a href="http://www3.gehealthcare.fr/"><img src="http://i.imgur.com/ryVz5vv.jpg" width="200" height="100" class="img-grey mautomargin"></a></li> | <li><a href="http://www3.gehealthcare.fr/"><img src="http://i.imgur.com/ryVz5vv.jpg" width="200" height="100" class="img-grey mautomargin"></a></li> | ||
<li><a href="https://www.corning.com/worldwide/en/products/life-sciences.html"><img src="http://i.imgur.com/ycgjpR7.jpg" width="225" height="100" class="img-grey mautomargin"></a></li> | <li><a href="https://www.corning.com/worldwide/en/products/life-sciences.html"><img src="http://i.imgur.com/ycgjpR7.jpg" width="225" height="100" class="img-grey mautomargin"></a></li> | ||
<li><a href="http://www.grandluminy.com/"><img src="http://i.imgur.com/GK5IMdz.jpg" width="200" height="100" class="img-grey mautomargin"></a></li> | <li><a href="http://www.grandluminy.com/"><img src="http://i.imgur.com/GK5IMdz.jpg" width="200" height="100" class="img-grey mautomargin"></a></li> | ||
| − | <li><a href="https://www.eppendorf.com/FR-fr/"><img src="http://i.imgur.com/jUjj2YD.jpg" width=" | + | <li><a href="https://www.eppendorf.com/FR-fr/"><img src="http://i.imgur.com/jUjj2YD.jpg" width="250" height="100" class="img-grey mautomargin"></a></li> |
<li><a href="http://www.dutscher.com/frontoffice/home;jsessionid=4164FFDE7B048361D89406E266542A13"><img src="http://i.imgur.com/wjZ6U1A.jpg" width="200" height="100" class="img-grey mautomargin"></a></li> | <li><a href="http://www.dutscher.com/frontoffice/home;jsessionid=4164FFDE7B048361D89406E266542A13"><img src="http://i.imgur.com/wjZ6U1A.jpg" width="200" height="100" class="img-grey mautomargin"></a></li> | ||
<li><a href="http://www.mn-net.com/"><img src="http://i.imgur.com/apuJ5in.jpg" width="200" height="100" class="img-grey mautomargin"></a></li> | <li><a href="http://www.mn-net.com/"><img src="http://i.imgur.com/apuJ5in.jpg" width="200" height="100" class="img-grey mautomargin"></a></li> | ||
Revision as of 06:20, 17 September 2015
Production of Laccase E.coli
To obtain our Laccase E.coli, we used the BioBrick Bba_K863006 and we removed its stop codon. Then we added to this BioBrick a promoter and a His-Tag.
Thanks to digestion, ligation and transformation, we managed to get our BioBrick named “01-35-02” with a size of about 1700 pb:
Figure A: Schematic representation of “01-35-02”
Figure A: Schematic representation of “01-35-02”
Figure B: Digestion from “01-35-02” miniprep to check the size of the insert
The band corresponds to the expected size for the insert, which is about 1700 pb.”
Figure C: Western Blot of the expression of “01-35-02” into E. coli
The expected size of the protein is about 53 kDa.
Laccase T.thermophilus from iGEM parts
To get our Laccase T. thermophilus, we used the BioBrick Bba_K863011 and we removed its stop codon.
Then we tried to add to this BioBrick a promoter and a His-Tag.
Unfortunately we managed to add only the promoter.
Figure A: Schematic representation of “01-35-02”
Then we inserted our BioBrick into E.coli strain (BL21) to express it. We induced it by addition of IPTG into the cell culture. We made a Western-Blot using a primary antibody against the His-tag and an anti-mouse secondary antibody conjugate with HRP (horseradish peroxidase). The expected size of the protein “01-35-02” is 53 kDa.
Then we tried to add to this BioBrick a promoter and a His-Tag.
Unfortunately we managed to add only the promoter.
Figure A: Schematic representation of “01-35-02”Then we inserted our BioBrick into E.coli strain (BL21) to express it. We induced it by addition of IPTG into the cell culture. We made a Western-Blot using a primary antibody against the His-tag and an anti-mouse secondary antibody conjugate with HRP (horseradish peroxidase). The expected size of the protein “01-35-02” is 53 kDa.
Figure E: Digestion from “01-36” miniprep to check the size of the insert
The band corresponds to the expected size for the insert, which is about 1500 pb.”Laccase T.thermophilus from IDT
We ordered from IDT a laccase optimized for an expression into E.coli.
From this optimised laccase, we added a promoter and a His-Tag.
This new BioBrick is named “01-30-02” with an expected size of about 1500 pb
Figure A : Schematic representation of “01-30-02”
From this optimised laccase, we added a promoter and a His-Tag.
This new BioBrick is named “01-30-02” with an expected size of about 1500 pb
Figure A : Schematic representation of “01-30-02”
Figure B: Digestion from “01-30-02” miniprep to check the size of the insert The band corresponds to the expected size for the insert, which is about 1500 pb.
