Cloning And Expression Ofgllal Cell Line－derived Neurotrophlc

Glial cell line-derived neurotrophic factor(GDNF) was initially purified and cloned from B49 glial cell line in 1993. Many studies show that GDNF has strong protection and restoration effect on rodent and primate models of neurodegenerative diseases. Recent study indicates that GDNF exerts both histologic and functional neuroprotective effects on rod photoreceptors in the rd/rd mouse. Furthermore, GDNF also play an important role in kidney development and spermatogenesis.Based on the GDNF gene sequence of human and chick, we cloned partial DNA sequences encoding mature protein of GDNF from genomic DNA of giant panda and crested ibis through PCR amplification. Sequence comparison showed the giant panda and crested ibis GDNF genes had high homology to counterparts of human, rhesus monkey, rat, mouse and chick. We constructed the pGEX-GDNF expression vecto'rs and obtained the recombinant protein of expected size after induction of IPTG. The fusion protein was purified using Glutathione Sepharose 4B affinity chromatography.