Figure 1

Expression and purification of the antiangiogenic (P60) and neuroprotective (P78) PEDF derivatives. (A) The N-terminal locations of P60 at residues 60–77 and P78 at residues 78–121 of the PEDF protein are indicated in the diagram. (B) Schematic showing cloning of the peptide derivatives into pET32a. Bands on the PCR gels represent the amplified P60 and P78 fragments. (C) On the SDS-PAGE gradient gel (4–20%) expression of P60 and P78 (U, uncleaved from the upstream thioredoxin vector sequence) and purity of the peptides after cleavage (C, cleaved) at 2.1 and 4.7 kDa, respectively, are shown (M, Bio-Rad Precision Plus Protein™ Dual Xtra Standards). (D) The Western blot confirms identity of the two fragments by using a PEDF polyclonal antibody. Calculated yields of the expressed (U) and purified (C) peptides are given in Table 1.