The present disclosure provides a recombinant adenovirus for expressing African swine fever virus (ASFV) EP153R-EP402R protein and a construction method thereof, and belongs to the technical field of genetic engineering. In the present disclosure, a recombinant adenovirus vector pAD-CMV-EGFP-EP153R-EP402R is obtained through a series of intermediate processes using a recombinant adenovirus shuttle vector pENTRE-EGFP-TOPO; the recombinant adenovirus vector is linearized to transfect AD293 cells, a recombinant virus is screened according to cytopathy formed by adenovirus infection, an adenovirus packaging process is achieved, and the recombinant adenovirus for expressing ASFV EP153R-EP402R protein is obtained, laying a foundation for the construction of a recombinant adenovirus vaccine for expressing the ASFV EP153R-EP402R protein.