Objective：This study aims to rescue a recombinant H1N1 pdm09 virus by reverse genetics method and evaluate its growth characteristics in MDCK cells and eggs as well as its pathogenicity in BALB/c mice. Methods：The six internal gene segments of A/Puerto Rico/8/1934（PR8）virus were amplified by RT-PCR and cloned into pHW2000 bidirectional expression vector. The sequences of hemagglutinin（HA）and neuraminidase（NA）of A/Wisconsin/588/2019（H1N1）were downloaded from global initiative of sharing all influenza data（GISAID）database，the HA non-coding region of the virus was replaced by that of A/California/07/2009 （H1N1）. HA and NA of A/Wisconsin/588/2019 were synthesized and cloned into pHW2000 bidirectional expression vector. Both the six internal gene recombinant plasmids of PR8 and the HA and NA recombinant plasmids of A/Wisconsin/588/2019 were co-transfected into 293T and MDCK cells to rescue the recombinant virus. The recombinant virus was identified by hemagglutination test and RT-PCR analysis. The recombinant virus was inoculated into MDCK cells and 9- 10 day-old eggs to evaluate their growth characteristics and draw growth curves. BALB/c mice were infected with the recombinant virus by intranasal drip. The changes in body weight and survival rate of the mice were recorded to evaluate the pathogenicity of the recombinant virus. Results：The recombinant A/ Wisconsin/588/2019 virus was rescued. Its hemagglutination titer was 1∶8，and the HA and NA sequences were consistent with expected results. Most MDCK cells were completely shed at 72 hours after inoculation with the recombinant virus，and its viral titer was 10^5.38TCID₅₀/mL. The peak of replication was 60 hours after inoculation. The hemagglutination titer reached 1∶2⁸ on eggs. All BALB/c mice died on the 8th day after intranasal administration of the recombinant virus with a dilution ratio of 1∶1，and the median lethal dose of the recombinant virus to mice was 10^-1.38/50 μL. Conclusion：A recombinant H1N1 pdm09 virus is successfully rescued. The virus grows well in MDCK cells and has good egg-adaptation. It is also pathogenic and lethal in mice. This study provides a new idea for the rapid rescue of influenza viruses by reverse genetics method.