Exchangeable design of regulatable AMPs and TAT-AMPs
CCU_Taiwan contributes the thrombin-releasable AMPs and TAT-AMPs in biobricks, and also provides flexibility for future modification.
The concept of biobrick allows novices to easily assemble the necessary components to express a protein. However, the prefix-suffix junction is translated into extra amino acids in the protein coding region, which may cause some problems in protein function. For example, a combination of thrombin site biobrick and AMP biobrick would leave two amino acids at the N-terminal of AMPs, hampering the AMP activity.
To deal with this functional issue of protein expression, we fused the thrombin site with AMPs or TAT-AMPs into a single biobrick, and further modified the peptide sequence in the thrombin site to optimize the AMP or TAT-AMP activity. ( You can find the details in design).
The numbers of AMP family members are still growing. We believe that there will be more powerful AMPs identified in the future. To provide the flexibility for exchanging AMPs in the future, we flank the thrombin-releasable AMPs and TAT-AMPs with two restriction enzyme sites, EcoRI and HindIII. These two restriction enzyme cutting sites will allow future iGEM teams to rapidly exchange the AMPs for a new one.