Part:BBa_K1800003:Experience

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Applications of BBa_K1800003

GSU iGEM 2016: Last year we performed a colony PCR on the part we sent in close to the 2015 Giant Jamboree. After the Giant Jamboree, we realized that we did not have the part in PSB1C3. This year we re-cloned it and confirmed that it was in PSB1C3 with sequencing.

The cDNA sequence for CBDA synthase was inserted into Agrobacterium tumefaciens using a binary vector system known as pORE. This allowed us to transfect tobacco plant tissue with agrobacterium that codes for CBDA synthase creating transgenic tobacco plants that will produce CBDA synthase in their roots. The roots of these plants will then be introduced to CBDA synthase substrate, Cannabigerolic acid, which will be catalyzed to form Cannabidiolic acid (CBDA). The Cannabidiolic acid will then be separated and heated at 120°C for 20min to form Cannabidiol. We plan to use the same transformation procedure to produce Horseradish Peroxidase (HRP) in the roots of transgenic tobacco, since it is more easily detected compared to CBDA synthase. One of the benefits of HRP is the bioluminescence that is produced when it catalyzes its substrate.

User Reviews

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