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Team:Frankfurt/Objectives
From 2012.igem.org
Results
Overexpression of GGPPS
Gap repair Cloning
Gap repair means the homologue recombination in yeast in order to assemble desired DNA fragments. It is easier and more effective than cloning via restriction and ligation. We used it to assemble the mevalonate overexpression plasmid and the steviol synthesis plasmid. All genes, promoters and terminators could be amplified except of the fragments for Gibson Assembly of KAH. Thus the mevalonate overexpression plasmid was assembled and transformed into yeast. To verify the correct assembly of the plasmid we did two control restrictions (see "Notebook"). Because we were not able to assemble KAH the gap repair of the steviol synthesis plasmid did not work. We also tried the gap repair with the four 500 bp fragments of KAH but the overlapping sequences are only 30 bp and not 40 bp which is needed for homologue recombination.
