Objective To explore the humoral and cellular immune responses of a vaccine of S fragments from a new type reovirus R4 strain in mice. Methods Four recombinant plasmids were constructed by respectively cloning S1, S2, S3,S4 genes into pcDNA3.1⁺, and mice were intramuscularly immunized with the recombinant plasmids in a dose of 100 μg/mouse. Control vector pcDNA3.1⁺ and phosphate buffered saline (PBS) were used as negative controls. The specific antibody level and IgG subclass (IgG1, IgG2a, and IgG2b) were detected by ELISA, and cellular immune responses to R4 were assessed using an interferon (IFN)-γ ELISpot assay. Results All recombinant plasmids induced significantly higher levels of anti-R4 IgG compared with that of the controls (pcDNA3.1⁺ and PBS), and the titers were highest in the mice immunized with S1 and S3. On the other hand, S1 gene induced highest IgG2a antibody and the cellular immune response was best. Conclusions After the mice immunized with S1 gene recombinant plasmid, this plasmid can initiate both cellular and humoral immune responses in mice. S1 gene recombinant plasmid is a promising vaccine candidate.