表达人CD40配体的重组腺病毒载体的构建

Objective To construct a recombinant adenovirus vector expressing hCD40Lgene and explore it in the use of anti- tumor gene therapy.Methods 1 900 bp gene fragmentwas obtained form plasmid pORF-hCD40L byXhoⅠ/SwaⅠcutting and then cloned directionally into the pShuttle plasmid, finally, the resultant plasmid was digested by restriction endonnucleasePmeI 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 digestionwere employed to identify the recombinant adenovirus.Results The evidences of endonulease digestion and PCR analysis confirmed that recombi- nant 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 tumorgene ther- apy.