Abstract: In order to construct a recombinant adenovirus type 5 expressing the novel Bunia virus（SFTSV） Gn gene, primers were designed and synthesized based on the SFTSV Gn gene sequence published in GenBank（ADZ04482.1）, and the KOZAK sequence and tPA signal peptide sequence were added to the Gn gene by specific PCR method. Then, a recombinant shuttle plasmid PGA-KOZAK-tPA-Gn was constructed by enzyme digestion and ligation, and homologous recombination with the adenovirus skeleton plasmid pAd5-ΔE1ΔE3-5E4 was performed in BJ5183 receptor cells to obtain the recombinant adenovirus plasmid rAd5-KOZAK-tPA-Gn. The recombinant adenovirus plasmid was cleaved using Pac Ⅰ enzyme, and the linearized plasmid was transfected into HEK 293 cells for virus packaging, propagation, and purification. PCR, Western-blot and other techniques were used to detect the expression of viral genes, and the titer of the recombinant adenovirus was determined. The results showed that 1 546 bp KOZAK-tPA-Gn gene was obtained by PCR amplification, and the recombinant shuttle plasmid was successfully constructed. SFTSV Gn gene was stable during the passage of recombinant adenovirus. Recombinant adenovirus expressed SFTSV Gn protein with molecular weight of 61 ku in HEK 293 cells. The titer of the recombinant adenovirus was measured to be 1×10^-7.63/0.1 mL TCID₅₀. These results indicated that recombinant adenovirus type 5 expressing SFTSV Gn gene was successfully constructed.