The experimental study of construction of recombinant plasmid pEGFP-GDNF and its expression in eukaryotic cell

Objective To construct a kind of engineered cells that can secrete human glial cell line derived neurotrophic factor(GDNF)and study its possible effects on genetic therapy of Parkinson's disease.Method The cDNA encoding the mature rat glial cell-derived neurotrophic factor (GDNF)was isolated was cloned into eucaryotic expression vector pEGFP-C1 of EGFP reported gene encoding green fluorescence protein in the form of fused protein.By using lipofectin and electric methods,It was transfected into SH-SY5Y cell line.Results The expression vector of recombinant plasmid pEGFP-GDNF was successfully constructed.Using fluorescence microscope analysis demonstrated that the GDNF gene can be transcripted and translated in SH-SY5Y cells.Conclusion It suggests that the engineered SH-SY5Y cells can secrete human GDNF and could play an important role in gene therapy of Parkinson's disease.