Construction Of Replication Defective Adenoviral Vector Carried SCD80-Linker-sCD40L Fusion Gene

Objective: To construct the replication defective adenoviral vector carried sCD80-Linker-sCD40L fusion gene. Methods: The coding gene of human sCD80, sCD40L and IL-12b signal were cloned by RT-PCR from human peripheral blood mononuclear cells (PBMC). We constructed IL-12bSig-sCD80-Linker-sCD40L fusion gene by overlap PCR, and used the AdEasy XL Adenoviral Vector System to generate replication defective adenoviral vector carried the sCD80-Linker-sCD40L fusion gene. Using westen blot to identify the fusion protein. Results: DNA sequence analysis proved that we cloned the coding gene of human sCD80, sCD40L and IL-12b signal, and we constructed the replication defective adenoviral vector carried sCD80-Linker-sCD40L fusion gene. Its liter is 2 1010cfu/ml. Westen blot shows that transfected 3T3 cells have secreted the fusion protein. Conclusion: We successful constructed the replication defective adenoviral vector carried sCD80-Linker-sCD40L fusion gene, which may be more helpful to further researches.