The invention discloses a preparation method of Burkholderia pseudomallei recombined BLF1 protein, the product prepared through the preparation method and an application of the Burkholderia pseudomallei recombined BLF1 protein. The preparation method includes the steps that a primer for amplifying a Burkholderia pseudomallei BLF1 gene is designed firstly, a sequence shown in SEQ ID NO.3 of a coding area sequence is obtained through amplification, then the acquired sequence is connected to an expression vector to construct a recombinant expression vector, the recombinant expression vector is converted into host bacteria, and through inducible expression, the Burkholderia pseudomallei recombined BLF1 protein is obtained. The preparation method is simple, the space design conception and immunogenicity of the purified protein can be kept to the maximum degree, the purity of the protein is larger than 95%, it is proved through a mouse experiment that the purified protein has animal toxicity, it is shown that the purified protein has biological activity, the purified protein acts on A549 cells, cell growth can be restrained, and thus the Burkholderia pseudomallei recombined BLF1 protein can be used for preparing anti-tumor medicine and has wide application prospects.