Human fgf2 cdna

MBP-FGF2 surfaces were prepared using our previously described method [38 (link)]. Briefly, MBP-FGF2 fusion proteins were obtained from Escherichia coli carrying pMAL-FGF2 plasmids that were generated by inserting human FGF2 cDNA (Bioneer, Korea) into pMAL vectors (New England Biolabs, U.K.). human FGF2 cDNA was cloned from human fibroblasts via polymerase chain reaction (PCR) using oligonucleotide pairs (5′-CCG AAT TCC CCG CCT TGC CCG AGG ATG GC-3′ and 5′-CAA AGC TTT CAG CTC TTA GCA GAC ATT GGA AG-3′; Bioneer) with EcoRI and HindIII restriction sites, respectively. The PCR products were cloned into pGEM-T plasmids (Promega, Madison, WI, USA) to generate pGEM-FGF2. pGEM-FGF2 and pMAL plasmids were digested using EcoRI-HindIII, recovered from an agarose gel, and ligated using a ligation kit (TaKaRa, Shiga, Japan) to generate pMAL-FGF2. MBP-FGF2 (20 μg/mL) spontaneously adsorbed onto polystyrene (PS) surface plates (non-tissue culture-treated 96 or 384 well plates; Falcon, Fisher Scientific, Forest Lawn, NJ, USA) at 37 °C for 4 h.