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. 2015 Oct 10;7(10):4067–4082. doi: 10.3390/toxins7104067

Figure 1.

Figure 1

Construction of the plasmid containing the dDTEGF13 gene. (A) The pET expression vector containing the dDTEGF13 target gene; (B,C) The PyMol sphere graphic was generated by downloading the Protein Data Bank [19] X-ray crystallographic structure of DT [20] into the PyMol 3D molecular modeling program [21] Shown is a frontal view of the protein and a 180° reverse view of the molecule; In (B), the amino acids associated with ADP-ribosylation (catalytic site) are bl ackened; In (C), The amino acids that were mutated for deimmunization are blackened; (D) SDS-PAGE gel analysis was performed to confirm the size and purity and stained with Coomasie blue. Photo is grayscale. Lane 1—Molecular weight standards, Lane 2—dDTEGF13 non-reduced, Lane 3—dDTEGF13 reduced. The gel was stained using Coomassie blue; (E) A HPLC trace for the purified drug is also shown illustrating mostly a single peak obtained from a TSK3000 size exclusion column. Only the single peak was collected resulting in a >95% purity.