Abstract: In the present study, researchers describe a rescue system for duck hepatitis virus-1 using T7 RNA polymerase. This strain can not induce typical cytopathic effect in BHK-21 cells. First, there is used the vitro transcripts to infect BHK-21 cells. Then the infected cell supernatant were inoculated into chicken embryos to rescue the virus and the cDNA clone was engineered to contain one silent nucleotide change to create a BssHII site to distinguish from the parental virus. The rescued virus was able to result in embryonic death and have similar infection with parental virus in chicken embryos indicating that the cDNA clones were replication competent. It is proposed that this rescue system may benefit the investigation on viral virulence determinants and molecular virology of DHV-1 virus.