Team:CSU Fort Collins/Improve
A New and Improved Lysostaphin Part
Our team needed a kill mechanism for our search and destroy circuit and after much research we found that lysostaphin was an excellent candidate. Thankfully, there was already a part designed by the 2012 HIT-Harbin team BBa_K748002 which we were able to grab and improve upon. We wanted to optimize the part for our vector species E. coli. The gene fragment was cloned into pET28a using the NcoI-XhoI site via Twist. To do this we started by codon optimizing the part for E. coli amino acid usage. Also a fusion protein was created with HlyA in order to increase secretion of lysostaphin. Next we edited out the cut sites and enterokinase sites from the original construction and added the Nco1 and Xho1 sites for synthesis in both the PlnE, PlnF, and lysostaphin +HlyA blocks.