Difference between revisions of "Team:Aachen/Lab/Methanol/Polycistronic Expression Plasmid"
Tschwanemann (Talk | contribs) (Created page with "{{Team:Aachen/Header}} __NOTOC__ In order to create a strain that is able to metabolize methanol, we wanted to build a polycistronic plasmid with all required genes for metha...") |
Tschwanemann (Talk | contribs) (→Results) |
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A SDS gel showed that all four genes are expressed. | A SDS gel showed that all four genes are expressed. | ||
− | Next, we tested the [[Team:Aachen/Lab/Methanol/Characterization | + | Next, we tested the [[Team:Aachen/Lab/Methanol/Characterization#Physiology|physiology]] and the [[Team:Aachen/Lab/Methanol/Characterization#Mdh_Characterization|enzyme activity ]] of a strain that carries our construct. |
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− | We intensively tried to clone the polycistronic construct into the pSB1C3 BioBrick backbone. After trials via [[Team:Aachen/Notebook/Protocols | + | We intensively tried to clone the polycistronic construct into the pSB1C3 BioBrick backbone. After trials via [[Team:Aachen/Notebook/Protocols#CPEC|CPEC]], restriction and ligation with different sets of enzymes, Gibson Assembly, and ligation after alkaline phosphatase digest, we decided to abandon this project. Our assumption is that a strain that carries the polycistronic expression frame in a BioBrick backbone is just not able to survive in a overnight culture, because clones from the LB plate after transformation never grew in a overnight culture. |
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'''[[Team:Aachen/Notebook/Documentation/Methanol_Polycistronic_Expression_Plasmid#Building_BioBricks_out_of_polycistronic_plasmid|Laboratory Notebook: Building BioBricks out of polycistronic plasmid]]''' | '''[[Team:Aachen/Notebook/Documentation/Methanol_Polycistronic_Expression_Plasmid#Building_BioBricks_out_of_polycistronic_plasmid|Laboratory Notebook: Building BioBricks out of polycistronic plasmid]]''' | ||
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=References= | =References= |
Revision as of 12:25, 18 September 2015