Aim To construct recombinant adenovirus vector containing human apoliproprotein AⅠ(apoAⅠ) gene and to explore the expression of human apoAⅠ in mice and establish human apoAⅠ overexpressing mice model. Methods Full length apoAⅠ cDNA was obtained from recombinant plasmid pDNR-LIB-apoAⅠ-PA via digestion with SalⅠ and HindⅢ and was subcloned into the pShuttle vector pshuttle CMV to construct a shuttle plasmid pshuttle CMV-AⅠ.Then it was co-transformed into E.coli BJ5183 with adenovirus genomic plasmid pAdEasy-1 to achieve homologous recombination pAd CMV-AⅠ.The DNA of identified recombinant plasmid was transfected into HEK293 cells via liposome mediation to package and amplify adenovirus AdV-AⅠ.BALB/c mice was administrated by purified adenovirus,then human apoAⅠ protein in the serum of mice was detected by double immunodiffusion,immunological turbidity kit and Western blotting. Results The titer of purified virus was 4.0 ×1013pfu/L.The over-expression of human apoAⅠ was observed after administration of BALB/c mice with adenovirus particles.Human apoAⅠ concentration in mice serum reached 740±110 mg/L at 1 day after administration of adenovirus particles. Conclusion The recombinant adenovirus AdV-AⅠ was successfully constructed and human apoAⅠ was detected in the serum of mice.The mice model of overexpressing human apoAⅠ was established.This result lays the foundation for further study to investigate the beneficial effects of apoAⅠ on endotoxemia.