The invention belongs to the field of animal genetic engineering, and relates to a ZB (zebrafish) transposon system of and a gene transfer method mediated by the ZB transposon system. The ZB transposon system is derived from zebra fish, and comprises two transgenic donor plasmids with terminal repetition sequences and capable of being inserted into a target gene box, and transposase auxiliary plasmids for providing activity of transposition; the transgenic donor plasmids comprise terminal repetition sequences and Msc1 insertion cloning sites at two sides of ZB transposon. The invention further discloses a gene transfer method based on the ZB transposon system, which is characterized that target genes are guided into a receptor genome through the ZB transposon system. The system and the method can be applied to multiple field of biotechnology: (1) the method can be used for effectively inserting the target gene box into a host cell genome, so that the gene transfer efficiency is improved; (2) by combining with the gene trapping technology, the researches on functions of animal genes can be effectively developed; (3) human gene therapy can be realized through mediation.