Team: iGEM Frankfurt -

Gap repair Cloning for Vector Assembly

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. This method was used to assemble the mevalonate overexpression plasmid in year 2012. Since in last year the KAH fragment, which is crucial for the second plasmid, was not able to be constructed the reconstruction of the steviol plasmid via gap repair was done this year. We were able to grow cultures containig the plasmid on selective media with the corresponding auxotrphy, but unfortunately we were to short on time to extract it.

Gas Chromatographic Analysis

Since the first plasmid created last year leads to a higher amount of geranylgeranylpyrophosphate, is was one of our aims for this year to prove that this actually worked out the way it was intended. For this reason we used gas chromatography coupled with mass spectrometry - but sadly didn't get the results in time to present them here.

iGEM-Team Frankfurt 2013