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From 2010.igem.org

Experimental results

generation of fusion proteins

The image shows all the fusion proteins that were planned. The BioBricks for all these constructs were generated via PCR or synthesized at GeneArt and ligated into the storage vector for sequencing. Easy cutting and pasting into working vectors should be possible with the cloning strategy BBF RFC 28.

Modeller's input

Due to the high amount of fusion proteins that were in planing (81 constructs), priorities had to be distributed to the different genes. This was possible due to the various models the dry-lab team implemented.

• Chemotactic protein: CheY was chosen as first target.
• Anchor: TetR was the first choice due to its wide application in synthetic biology and extensive characterization.
• Pif3 linked to Che-Protein: Although PhyB seemed to be the more robust choice, Pif3 was chosen to be linked to the Che-Protein due to the tendency of PhyB to sequester.
• Ratio anchor (i.e. tetO) to binding partner (i.e. tetR): The simulations favoured a ratio of 50 µM anchor to 40 µM anchor binding partner.

Modeller's input

Due to the high amount of fusion proteins that were in planing, priorities had to be distributed to the different genes. This was possible due to the various models the dry-lab team implemented.