The invention discloses a method for preparing an African swine fever virus P30 and P54 yeast vaccine. The vaccine contains African swine fever virus strong immunogen protein P30, membrane structure protein P54, Fc fragments of swine IgG1 and IgA1 (namely Fc gamma, Fc alpha), and His purification tags. The method comprises the following steps: S1, connecting ASFV immunogen protein genes p30 and p54 with Fc gamma and Fc alpha in series; S2, respectively constructing transcription units of p30-Fc gamma and p54-Fc alpha in fusion expression in the yeast; S3, realizing series connection of the twotranscription units through in-vitro enzyme linking, yeast in-vivo conversion and homologous recombination technologies, and stably integrating the two transcription units into surface display type saccharomyces cerevisiae; and S4, carrying out genome level and protein level verification, and confirming to obtain two recombinant yeast strains of which the independent surfaces show and express theP30-Fc gamma and P54-Fc alpha fusion proteins, and a tandem composite expression type recombinant yeast strain of which the surface shows and expresses the two fusion proteins at the same time in S1.The strain is mature in production technology, suitable for large-scale production, safe to use and capable of inducing remarkable mucous membrane and humoral immune response of target animals.