Construction Of Recombinatant Adenovirus Carrying GDNF And EDNRB Gene And It's Coexpression In Neural Stem Cells

Objective To construct the recombinant adenovirus bearing glialcellline derived neurotrophic factor (GDNF) and endothelin receptor B (EDNRB) gene and investigate its coexpression in neural stem cells. Method The AdEasy system was used to gene vate the recombinant Adenovirus Ad-GE coexpressing GDNF and EDNRB gene . The expression of exogenous genes in neural stem cells after infection was determine serving the expression of the green fluorescence protein(GFP) and detected by reverse transcription polymerrase chain reaction (RT-PCR) method and real-time fluorogenic quantitative PCR. Result GFP expression could be observed under fluorescent microscope 24h later after infection and coexpression of GDNF and EDNRB gene was confirmed by RT-PCR method and real-time fluorogenic quantitative PCR. Conclusion Exogenous GDNF and EDNRB gene can be coexpressed in neural stem cells , which may offer the possibility of a novel approach to local combination gene therapy of Hirschsprung disease .