Objective To establish the sperm specific Sleeping Beauty (SB) transposase-expression transgenic mouse for the study of the genetic modification mediated by transposon system in mouse. Methods Prm1 promoter was cloned from mouse genomic DNA to drive the expression of SB transposase. The transgenic mice were generated by microinjection. The gene type of transgenic line was identified by PCR. The expressing level in testis was determined by western blot and immunohistochemistry (IHC) staining. Results Five lines of transposase transgenic mice were obtained by microinjection and three can be germline. One mouse line with higher expression level of transposase in the testis was obtained. Conclusion One transgenic mouse model with Sleeping Beauty transposase-expression was successfully established. This model will greatly contribute to the research of genetic modification mediated by transposon in mouse