Construction of the recombinant adenovirus vector expressing human CD40 ligand gene

Objective To construct a recombinant adenovirus vector expressing hCD40L gene and explore it in the use of antitumor gene therapy.Methods 1 900 bp gene fragment was obtained form plasmid pORF-hCD40L by XhoⅠ/SwaⅠ cutting and then cloned directionally into the pShuttle plasmid,finally,the resultant plasmid was digested by restriction endonnuclease PmeI and subsequently cotransformtion into BJ5183 cells with the adenoviral backbone pAdEasy-1 to obtain the homologous recombinant and then the recombinant was packaged in the 293 cells.Some methods such as PCR and endonulease digestion were employed to identify the recombinant adenovirus.Results The evidences of endonulease digestion and PCR analysis confirmed that recombinant hCD40L gene was correctly inserted into adenovirus vector.Conclusion The adenoviral vector which expressed hCD40L gene was constructed.It provides an experimental basis for studies on it expression in the mammalian cells and in tumor gene therapy.