Difference between revisions of "Team:Aix-Marseille/Results"
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| − | <h2 class="title wow Hinge"><span style="color:#8E3B8C"> | + | <h2 class="title wow Hinge"><span style="color:#8E3B8C">Production of Laccase E.coli</h2></span> |
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| − | <p class="space20"><div align="justify"> | + | <p class="space20"><div align="justify"> |
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| + | 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: <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 /> | ||
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| + | </div></p> | ||
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| − | <h2 class="title wow Hinge"><span style ="color:#000000"> | + | <h2 class="title wow Hinge"><span style ="color:#000000">Laccase T. thermophilus</span></h2> |
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| − | <p class="space20"><div align="justify"><span style ="color:#000000"> | + | <p class="space20"><div align="justify"><span style ="color:#000000"> |
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| + | • Laccase T. thermophilus from iGEM parts<br /><br /> | ||
| + | |||
| + | 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 /><br /> | ||
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| + | • Laccase T. thermophilus from IDT<br /><br /> | ||
| + | |||
| + | 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 /> | ||
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| + | </p></span></div> | ||
<p><div align="justify"><span style ="color:#000000">TEXTE</p></span></div> | <p><div align="justify"><span style ="color:#000000">TEXTE</p></span></div> | ||
<p><div align="justify"><span style ="color:#000000">TEXTE</p></span></div> | <p><div align="justify"><span style ="color:#000000">TEXTE</p></span></div> | ||
Revision as of 17:16, 15 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:
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 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.
Laccase T. thermophilus
• 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.
• 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.
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.
• 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.
TEXTE
TEXTE
TEXTE
TEXTE
