Design of constructs for the expression and purification of Drosophila melanogaster morphogens

The experiments were performed at the Center for Medicinal Chemistry (CQMED), which applies SGC protein purification strategies for drug-discovery approaches. These strategies involve generating different constructs of aim proteins and testing a small-scale high-throughput assay to identify the best constructs for generating large-scale aim proteins.

The Center for Medicinal Chemistry (CQMED), which is part of the SGC, is where the experiments were performed. This work involved four variants of each morphogen protein, with the following variables: full length; truncated at the N-terminal; truncated at the C-terminal; or both truncated. Each of these four variants was cloned into five different plasmids using Ligation Independent Cloning (LIC): pNic28-Bsa4 (adds a His-tag to the N-terminal region, linked by a TEV cleavage site), pGTVL2 (adds a His-tag and a GST tag to the N-terminal region, linked by a TEV cleavage site), pNIC-CTH0 (adds a His-tag to the C-terminal region, linked by a TEV cleavage site), pNIC-Zb (adds a His-tag and a Z-basic tag (ZB) to the N-terminal region, linked by a TEV cleavage site), and pSUMO-LIC (adds a His-tag and a SUMO protein).