Complete nucleotide sequence of a novel porcine circo- virus-like agent and its infectivity in vitro

A novel agent (hence termed as P2) was isolated from pig sera in China, which contained covalently bound circular genomic DNAs of 993 nucleotides. Sequence analyses indicated that the agent was closely related to the porcine circovirus (PCV). The molecular clone of P2 was constructed subsequently and used for the following studies. Intracytoplasmic inclusions and intranuclear inclusions were only found in PK-15 cells transfected with the tandem dimer of P2 molecular DNA clone. Intracytoplasmic inclusions were round or irregular in shape and 0.1–0.4 μm in diameter, and intranuclear inclusions were electronically denser than intracytoplasmic inclusions and had two general shapes: round/small (0.1 μm in diameter) and hexagonal/large (0.5–1.4 μm in diameter). The inclusions were not membranously bound. The cells transfected with the tandem dimer of P2 molecular DNA clone were tested positive for P2 DNA at passages 5. The P2 antigen could be detected in both transfected and passaged PK-15 cells. This is the first report regarding the complete nucleotide sequence of a small DNA genome in a circovirus-like infectious agent in vitro. Supported by the State Key Basic Research Project (973 project) of China (Grant No. 2007CB116308), Planned Projects for Postdoctoral Research Funds of Jiangsu Province, China (Grant No. 5910602) and Postdoctoral Funds of Jiangsu Academy of Agricultural Sciences, China (Grant No. 6510501)