Abstract:

The aim of this study was to construct an infectious DNA clone of porcine circovirus type II (PCV2) and lay the foundations for the mutant construction and pathogenesis researches of the PCV2. The complete genome of PCV2 was amplified from the PCV2 DNA plasmid by PCR method, and inserted into the pBluescript-SK vector to construct the single copy of PCV2 DNA clone. Furthermore, the double copy of PCV2 DNA clone was developed. Then, the transfection of porcine kidney (PK)-15 cells with the PCV2 DNA clones was conducted to obtain the rescue viruses of PCV2, and the reproducibility of the rescue virus in vitro was determined. The results showed that the rescue viruses of PCV2 were acquired successfully and could propagate stably in PK-15 cells, its 50% tissue culture infectious doses (TCID50) was 10?6.05/mL after 5 serial passages, which was close to its parent virus and demonstrated its relatively high infectious titer. The research results revealed that an infectious clone of PCV2 with relatively high infectivity was developed successfully, which established a foundation for further research on the virus biological characteristics and pathogenesis.