To obtain active hFKBP52 protein for screening novel neurotrophic drugs. Semi-nested and overlap PCR and affinity chromatography were used. hFKBP52 gene was cloned successfully from human fetal brain cDNA library, and then highly expressed (about 30%) as fusion protein in pET28a(+) vector system. The recombinant protein was purified as one band on SDS-PAGE. The purified hFKBP52 showed peptidyl-prolyl cis-trans isomerase (PPIase) activity, similar to the wild type.