[Construction of eukaryotic expression vector for human glial derived neurotrophic factor and its expression in spinal cord tissue of SD rat].
OBJECTIVE
To investigate the possibility of constructing eukaryotic expression vector for human glial derived neurotrophic factor (hGDNF), transfecting it to spinal cord tissue of rats so as to treat acute spinal cord injury.


METHODS
The eukaryotic expression vector pcDNA3-hGDNF was constructed by recombinant DNA technique, transfected into glial cell and neuron of spinal cord by liposome DOTAP as experimental group. In control group, mixture of empty vector and liposome was injected. The mRNA and protein expressions of hGNDF were detected by RT-PCR and Western blot.


RESULTS
After the recombinant eukaryotic expression vector for hGDNF was digested with Hind III and Xba-I, electrophoresis revealed 400 bp fragment for hGDNF gene and 5 400 bp fragment for pcDNA3 vector. In the transfected spinal cord tissue, the mRNA and protein expressions of hGDNF gene were detected with RT-PCR and Western blot.


CONCLUSIONS
The constructed eukaryotic expression vector pcDNA3-hGDNF could be expressed in the transfected spinal cord tissue of rat, so it provide basis for gene therapy of acute spinal cord injury.
Li
Qi
Q
Department of Orthopedics, the Sixth People's Hospital, Shanghai Jiaotong University, Shanghai, 200233, P. R. China.
Zeng
Bingfang
B
Xu
Jianguang
J
D000818:Animals; D005260:Female; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D008297:Male; D010957:Plasmids / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013116:Spinal Cord / Q000378:metabolism*; D013119:Spinal Cord Injuries / Q000628:therapy*; D014162:Transfection
D051100:Glial Cell Line-Derived Neurotrophic Factor; D012333:RNA, Messenger