Related Experiment Video
Updated: Jul 14, 2026
Adeno-associated Virus-mediated Transgene Expression in Genetically Defined Neurons of the Spinal Cord
Published on: May 12, 2018
[Construction of eukaryotic expression vector for human glial derived neurotrophic factor and its expression in
Qi Li1, Bingfang Zeng, Jianguang Xu
1Department of Orthopedics, the Sixth People's Hospital, Shanghai Jiaotong University, Shanghai, 200233, P. R. China.
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.
Conclusion:
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.
