Team:GeorgiaTech/Parts
Team Parts
Parts Table
"BioBricking"
In order to assemble our BioBricks, we chose the protein plastocyanin (PDB: 1JFX) to clone into the pSB1C3 vector. We performed 24 separate error-prone PCR reactions on plastocyanin as described on the Experiments page, and ligated the error-amplified plastocyanins into the pSB1C3 vector. We verified that our amplified samples were indeed ligated by running gel electrophoresis.
We confirmed that 4 of the samples were correctly ligated, and sequenced these four samples in order to align them.
The DNA alignment to the right yields a calculated error-rate of 0.2% - 0.5%, which was the expected rate of error from our previous experience. Success! The four plastocyanin mutants, along with the plastocyanin wild-type, were submitted to the Registry.
These BioBricks will be available for all future iGEM teams to use. In particular, we hope that Georgia Tech's iGEM team can continue our project next summer by expanding our mutation library further and performing screening tests! Our efforts have laid a solid foundation for future work in improving the CuAAC reaction and we are optimistic that in the coming years, the CuAAC reaction will eventually find roles in living systems.