Construction of infectious molecular clone of Duck hepatitis virus type 1 CL strain

Abstract:

Abstract: [Objective] To construct an infectious clone for studying functions of duck hepatitis virus(DHV) type1 genome by reverse genetic technique. [Methods] Three fidelity DNA fragments covering the full genome of DHV type1 CL strain were amplified by RT-PCR, and inserted into pBR322 vector, resulting in the full-length cDNA clone BR-CL. The in vitro-transcribed RNA from BR-CL was transfected into duck embryo renal cells and the rescued virus was identified using RT-PCR, indirect immunofluorescence assay and colloidal gold immunoelectron microscopy after six generations. After inoculating the rescued virus into SPF chick embryos, embryo death and pathological changes were observed. [Results] The results of RT-PCR, indirect immunofluorescence assay and immunoelectron microscope showed that infectious virus was rescued. After inoculating into SPF chick embryos, the rescued virus was able to kill embryos with pathogenic changes. [Conclusion] This is the first report on generation of infectious cDNA clone of DHV, which provides a valuable platform for further research on functions of DHV genome.