Difference between revisions of "Team:Aix-Marseille/Results"
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Thanks to digestion, ligation and transformation, we managed to get our BioBrick named “01-35-02” with a size of about 1700 pb: <br /><br /> | Thanks to digestion, ligation and transformation, we managed to get our BioBrick named “01-35-02” with a size of about 1700 pb: <br /><br /> | ||
| − | + | <img src=https://static.igem.org/mediawiki/2015/7/71/Figure_A.png class="img-responsive" width="400" height="250"> | |
| − | + | <span>Figure A: Schematic representation of “01-35-02”</span><br /><br /> | |
| − | + | ||
</div></p> | </div></p> | ||
</div> | </div> | ||
| + | |||
| + | |||
<div class="col-md-5 col-md-offset-1 col-sm-offset-1 space30"> | <div class="col-md-5 col-md-offset-1 col-sm-offset-1 space30"> | ||
| − | + | ||
| + | <div class="success-work project"> | ||
<div class="success-work-desc"> | <div class="success-work-desc"> | ||
| − | <img src= | + | <img src= https://static.igem.org/mediawiki/2015/7/7f/Figure_B.png class="img-responsive" width="180" height="113"> |
| − | + | <span>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.”</span><br /><br /> | |
| + | |||
</div> | </div> | ||
</div> | </div> | ||
| − | + | <div class="success-work project"> | |
| + | <div class="success-work-desc"> | ||
| + | <img src= https://static.igem.org/mediawiki/2015/c/c1/Figure_C.png class="img-responsive" width="250" height="156"> | ||
| + | <span>Figure C: Western Blot of the expression of “01-35-02” into E. coli | ||
| + | The expected size of the protein is about 53 kDa. </span> | ||
| + | |||
| + | |||
</div> | </div> | ||
| + | </div> | ||
</div> | </div> | ||
<div class="space30"></div> | <div class="space30"></div> | ||
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<!-- start science section --> | <!-- start science section --> | ||
| − | <section style="padding:30px 0px 50px | + | ------------------------------ |
| − | + | ||
| + | <div class="clearfix"></div> | ||
| + | <!-- start project section --> | ||
| + | <section style="padding:30px 0px 50px;" class="arrow_box" id="team"> | ||
| + | |||
<div class="container"> | <div class="container"> | ||
<div class="row"> | <div class="row"> | ||
| − | |||
<div class="col-md-6 left"> | <div class="col-md-6 left"> | ||
| − | <h2 class="title wow Hinge"><span style ="color:# | + | <h2 class="title wow Hinge"><span style="color:#8E3B8C">Laccase <i>T.thermophilus</i> from iGEM parts</h2></span> |
<div class="space30"></div> | <div class="space30"></div> | ||
| − | <p class="space20"><div align="justify | + | <p class="space20"><div align="justify"> |
| − | + | To get our Laccase T. thermophilus, we used the BioBrick Bba_K863011 and we removed its stop codon. <br />Then we tried to add to this BioBrick a promoter and a His-Tag.<br /> Unfortunately we managed to add only the promoter.<br /> | |
| − | + | <img src= https://static.igem.org/mediawiki/2015/f/fe/Figure_D.png class="img-responsive" width="400" height="250"> | |
| + | <span>Figure A: Schematic representation of “01-35-02”</span><br /><br /> | ||
| − | + | 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. <br /> | |
| − | + | </div></p> | |
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
</div> | </div> | ||
| − | <div class="col-md-5 col-md-offset-1 col-sm-offset-1 space30 | + | |
| − | + | ||
| + | <div class="col-md-5 col-md-offset-1 col-sm-offset-1 space30"> | ||
| + | |||
| + | <div class="success-work project"> | ||
<div class="success-work-desc"> | <div class="success-work-desc"> | ||
| − | + | <img src= https://static.igem.org/mediawiki/2015/4/43/Figure_E.png class="img-responsive" width="400" height="250"> | |
| − | + | <span>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.”</span><br /><br /> | |
| + | |||
</div> | </div> | ||
| + | |||
</div> | </div> | ||
| − | + | </div> | |
| + | <div class="space30"></div> | ||
| + | </div> | ||
| + | |||
| + | </section> | ||
| + | <div class="clearfix"></div> | ||
| + | ------------------------------ | ||
| + | |||
| + | <div class="clearfix"></div> | ||
| + | <!-- start project section --> | ||
| + | <section style="padding:30px 0px 50px;" class="arrow_box" id="team"> | ||
| + | |||
| + | <div class="container"> | ||
| + | <div class="row"> | ||
| + | <div class="col-md-6 left"> | ||
| + | <h2 class="title wow Hinge"><span style="color:#8E3B8C">Laccase <i>T.thermophilus</i> from IDT</h2></span> | ||
| + | <div class="space30"></div> | ||
| + | <p class="space20"><div align="justify"> | ||
| + | |||
| + | We ordered from IDT a laccase optimized for an expression into E. coli.<br /> From this optimised laccase, we added a promoter and a His-Tag.<br /> This new BioBrick is named “01-30-02” with an expected size of about 1500 pb <br /><br /> | ||
| + | <img src="https://static.igem.org/mediawiki/2015/a/a8/Figure_A2.png" width="500" height="179" space="0"> | ||
| + | <span>Figure A : Schematic representation of “01-30-02”</span><br /><br /> | ||
| + | </div></p> | ||
| + | |||
| + | |||
</div> | </div> | ||
| + | |||
| + | |||
| + | <div class="col-md-5 col-md-offset-1 col-sm-offset-1 space30"> | ||
| + | |||
| + | <div class="success-work project"> | ||
| + | <div class="success-work-desc"> | ||
| + | <img src="https://static.igem.org/mediawiki/2015/5/58/Figure_B2.png" space200> <br /> | ||
| + | <span>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.</span><br /><br /> | ||
| + | |||
| + | </div> | ||
| + | |||
| + | </div> | ||
| + | |||
</div> | </div> | ||
| + | <div class="space30"></div> | ||
</div> | </div> | ||
</section> | </section> | ||
| − | + | <div class="clearfix"></div> | |
| + | |||
</section> | </section> | ||
Revision as of 16:44, 16 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.
