Objective: To construct a recombinant plasmid including the gene coding the major allergen of Platanus acerifolia pollen. Methods: Total RNA was extracted from natural Platanus acerifolia pollen and cDNA was revese-transcripted. The DNA sequence of major allergen of platanus acerifolia pollen was amplified by polymerase chain reaction（PCR）. The linkage between amplified DNA sequences and plasmid pET32a was realized by double enzyme digestion. The recombinant plasmid was subsequently transferred into E.coli DH5α to prepare gene clone by CaCl2 method. The cloned recombinant plasmid was extracted by alkaline lysis method and the target fragment was confirmed by agarose electrophoresis， followed by PCR and sequencing. Results: The recombinant plasmid including the gene coding the major allergen of Platanus acerifolia pollen was constructed. Conclusion: The recombinant plasmid which included the gene coding the major allergen could be successfully constructed by using natural Platanus acerifolia pollen, and will be available for inducing expression of the recombinant major allergen protein.