Objective: To provide a highly efficient adenoviralvector Ad-CMV-hTGFβ1 for the study of gene therapy forreversion of the intervertebral disc degeneration.Methods: A newly developed recombinant adenoviralvector construction system was used in the study. ThecDNA of hTGFβ1 was first subcloned into a shuttle plasmidpShuttle-CMV. The resultant plasmid was linearized bydigesting with restriction endonuclease PmeI, andsubsequently transformed into E.coll. BJ5183 cells with anadenoviral backbone plasmid pAdEasy-1. Recombinantswere selected by kanamycin resistance and confirmed byrestriction endonuclease analysis. Finally, the recombinantplasmid linearized by PmeI was transfected into 293 cells.Recombinant adenoviruses were generated within 2 weeks.Results: The recombinant adenoviral plasmids werecut by BamHI and PacI respectively, and the diagnosticfragments appeared in 0.8% agarose electrophoresis. Theinfected 293 cells showed evident cytopathlc effect (CPE).The productions of PCR confirmed the presence ofrecombinant adenovirus. The expression of hTGFβ1 wasverified by immunohistochemical staining.Conclusions: The successful generation of theadenoviral vector Ad-CMV-hTGFβ1 and the confirmationof the interest gene expression make it possible for theexperimental study of the reversion of the intervertebraldisc degeneration by gene therapy.