Abstract: Objective: To study the feasibility and effect of gene therapy for murine melanoma by replication-defective recombinant adenovirus-induced mIFN-βgene. Methods: The gene of murine interferon beta (mIFN-β ) was transduced into murine melanoma cells B16 by human replicationdefective recombinant adenovirus via ex vivo and in vivo respectively, then the tumorigenicity, tumor vaccine effect and anti-tumor effect were investigated. Results: 1) The recombinant adenoviruses harboring foreign gene infecting B16 cells had high efficiency of gene transfer and expression; 2) The transfection of AdmIFN- βhad no effect on the growth rate of B16 cells, but the expression of major histocompatibility complex class (MHC-I) of those cells was obviously enhanced; 3) In mouse experiments, ex vivo mIFN-βgene delivered into B16 cells by adenovirus not only resulted in an efficient inhibition of tumorigenicity in situ, but also led to a growth inhibition of parental cells at other sites; 4) Giving AdCMVmIFN-βthrough i.t. and i.v resulted in a significant anti-tumor effect on subcutaneous tumors and lung metastasis. Conclusion: It' s feasible and effective of gene therapy for murine melanoma by replication-defective recombinant adenovirus harboring mIFN-βgene. This study indicates it has bright prospect to develop tumor vaccine and treat tumor by using adenovirus vector harboring a certain gene.