Difference between revisions of "Team:Aachen/InteractiveTour52"

Revision as of 23:24, 18 September 2015

To implement the MCC, we successfully built and characterized a polycistronic plasmid expressing all four MCC genes. With this construct we modified an E. coli strain so that it can tolerate higher methanol concentrations. Furthermore, we proved the functional heterologous expression of the methanol dehydrogenase which we considered to be the bottleneck of this pathway.

In parallel to establishing methanol as a carbon source for E. coli, we also enabled it to accumulate remarkable amounts of glycogen. We created and characterized single knockouts of glgX and glgP in Escherichia coli BL21 Gold (DE3). This was done with CRISPR/Cas9 genome editing which we found to be a very efficient method for genome modifications. In addition we assembled and characterized a functional glycogen synthesis operon.

By combining the characterized synthesis operon (glgCAB) and the knockout of glgP in one organism, further enhanced glycogen formation.

Finally, we fused our two modules by introducing the methanol operon into our BL21 Gold (DE3) ΔglgP strain.

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 <p class="headline" style="display:none;">Biology</p>
-Both modules of the biological approach
-To implement the MCC, we successfully built and characterized a polycistronic plasmid expressing all four MCC genes. With this construct we modified an E. coli strain so that it can tolerate higher methanol concentrations.
-Characterizing the functional expression of Bacillus methanolicus methanol dehydrogenase 2 in E. coli
-Developing an efficient cloning strategy to build a monocistronic diversity library using the RDP standard
+To implement the MCC, we successfully built and characterized a polycistronic plasmid expressing all four MCC genes. With this construct we modified an ''E.&nbsp;coli'' strain so that it can tolerate higher methanol concentrations. Furthermore, we proved the functional heterologous expression of the methanol dehydrogenase which we considered to be the bottleneck of this pathway.
-Created and characterized single knockouts of glgX and glgP in Escherichia coli BL21 Gold (DE3)
+In parallel to establishing methanol as a carbon source for ''E.&nbsp;coli'', we also enabled it to accumulate remarkable amounts of glycogen. We created and characterized single knockouts of glgX and glgP in Escherichia coli BL21 Gold (DE3). This was done with CRISPR/Cas9 genome editing which we found to be a very efficient method for genome modifications. In addition we assembled and characterized a functional glycogen synthesis operon.
-Achieved a double knockout ∆glgP/glgX in E.coli NEB10β
-Assembled and characterized a functional glycogen synthesis operon
+By combining the characterized synthesis operon (''glgCAB'') and the knockout of glgP in one organism, further enhanced glycogen formation.
-Combined and characterized synthesis operon (glgCAB) and knockout of glgP in one organism
+Finally, we fused our two modules by introducing the methanol operon into our BL21 Gold (DE3) Δ''glgP'' strain.
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