Experiments
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Scaffold
Enzymes
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Laboratory Objective #1 - Produce the Assemblase Scaffold
This is the first of our three laboratory objectives. We aim to express and purify βPFD-SnoopCatcher and αPFD-SpyCatcher. The assembly of these proteins into the Assemblase complex will then be demonstrated. Conjugation of SpyTag to SpyCatcher and SnoopTag to SnoopCatcher will also be demonstrated. To learn more see our Assemblase Scaffold page.
Laboratory Objective #2 - Produce and test PAM and TycA of Pathway 1
Our second lab objective was to assemble and test pathway 1. We aimed to express and purify the reaction enzymes involved in the biosynthesis of the Paclitaxel side-chain; PAM-SnoopTag and TycA-SpyTag. These enzymes will then be conjugated to βPFD-SnoopCatcher and αPFD-SpyCatcher, respectively. The catalytic activity of the enzymes will also be tested. To learn more see our Pathway 1 page.
Laboratory Objective #3 - Produce and test LXYL and DBAT of Pathway 2
This is the third of our three laboratory objectives. We aim to express and purify the reaction enzymes involved in the alternative biosynthesis of Paclitaxel; DBAT-SnoopTag and LXYL-SpyTag. These enzymes will then be conjugated to βPFD-SnoopCatcher and αPFD-SpyCatcher, respectively. The catalytic activity of the enzymes will also be tested. To learn more see our Pathway 2 page.