Figure 8
From: Functional characterization of an ornithine cyclodeaminase-like protein of Arabidopsis thaliana
Expression and purification of recombinant AtOCD. A. Commasie blue stained SDS-PAGE gel showing different purification steps of GST fused AtOCD and western blot using anti GST antibody to confirm the identity of the GST-AtOCD band. B. Time course analysis with purified AtOCD protein showed no change in the absorbance reading after 30 min of incubation with the putative substrate ornithine and NAD+. Data are representative results of one out of 3–4 independent experiments using independent purifications of recombinant AtOCD. Data are means ± SE (N = 2-3) of replicate assay tubes. Similar results were obtained either from bacteria or plant purified AtOCD protein and using different substrate and cofactor combinations described in the text. C. AtOCD fused with FLAG immunoprecipitated from T3 transgenic plants and detected by western blot. A mock purification from wild type plants was used as a control. The band migrated at the expected size of AtOCD:Flag (37.7 kD).