The Human Recombinant GDNF and Study of
Its Biological Activity
CHEN Zhe-Yu, CHEN Xiu-Qing and LU Chang-Lin
(Department of Neurobiology, the Second Military Medical University, Shanghai
200433, China)
SUN Jian-Xin and WU Xiang-Fu
(Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai
200031, China)
Abstract The cDNA encoding the mature
human glial cell derived neurotrophic factor(GDNF) was isolated by
using RT-PCR method from total RNA extracted of fetal human brain. The
expression plasmid pET-GDNF was constructed by inserting GDNF
cDNA into plasmid pET-28a(+) containing T7 promoter and transformed into E.coli
BL21(DE3). An expression strain BLGDNF was selected. SDS-PAGE analysis
revealed that the human GDNF protein was highly expressed and accumulated up to
above 30% of the total bacterial proteins in the form of inclusion body after
the induction. The antibody to GDNF was prepared by immunization of
rat using purified GDNF protein. Purified and refolded GDNF protein
could significantly promote the survival of dopaminergic neurons.
Key Words GDNF; gene cloning; gene
expression; dopaminergic neuron
Corresponding author:
Tel, 86-21-64374430-292; Fax, 86-21-64338357; e-mail, [email protected]