Team:Bielefeld-Germany/Amsterdam/Results

From 2012.igem.org

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<h1>Subtrate Analysis</h1>
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<h1>Substrate Analysis</h1>
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Since Amsterdam team activity test, team immobilization and team substrate analysis used laccases from the same pool.
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After the regionals Team Activity Test, Team Immobilization and Team Substrate Analysis got laccases from the same pool from the Cultivation Team. With these laccases the degradation experiments of estradiol and ethinyl estradiol were repeated for BPUL and ECOL. The new laccases BHAL and TTHL were characterized for estradiol and ethinyl estradiol degradation, too. <p class="more"> For more information about the new results[https://2012.igem.org/Team:Bielefeld-Germany/Results/substrate#Degradation_of_estrogens click here].
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With these laccases we repeated the degradation experiments. For BPUL and ECOL more biological replicates were measured and the laccases BHAL and TTHL were characterized for estradiol and ethinyl estradiol degradation. Possible resulting degradation products were futher analysed via LCMS-MS.
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For more informations [https://2012.igem.org/Team:Bielefeld-Germany/Results/substrate click here]
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Possible resulting degradation products were further analysed via LCMS-MS.
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Revision as of 19:26, 26 October 2012

Results since Regionals

Summary

coming soon

Laccases

The iGEM Team successfully produced again four active bacterial laccases:

All of the bacterial laccases were accomplished to purify. The purified laccases . Furthermore the iGEM Team Bielefeld demonstrated that the produced laccases can be immobilized maintaining their activity and the degradation capacity was screened for several micro-contaminants. These tests indicate that the ECOL and BPUL are able to degrade ethinyl estradiol and estradiol. The laccase of TVEL0 fromTrametes versicolor was produced and tested for activity on oaxidation of ABTS.

Immobilization

Coming soon!

For immobilization results see here

Substrate Analysis

After the regionals Team Activity Test, Team Immobilization and Team Substrate Analysis got laccases from the same pool from the Cultivation Team. With these laccases the degradation experiments of estradiol and ethinyl estradiol were repeated for BPUL and ECOL. The new laccases BHAL and TTHL were characterized for estradiol and ethinyl estradiol degradation, too.

For more information about the new resultsclick here.

Possible resulting degradation products were further analysed via LCMS-MS.


Cellulose binding domain

A lot of efforts were made, to change the order of the fusion proteins, to change the promoter and the RBS and to insert a different linker between the cellulose binding domain and the reporter GFP, but to the last day of lab work no green glowing colony to work with and execute the binding assay could be generated.

Read '''the Boston file'''

Shuttle vector

Coming soon! Read more.


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