The largest open reading frame of porcine circovirus (ORF 4) encodes a protein of 312 amino acids. The predicted gene product of ORF 4 shows similarities to Rep proteins of other plant circoviruses and geminiviruses. Three motifs have been identified that are characteristic for proteins involved in rolling circle replication and the consensus sequence for a putative dNTP-binding box (GKS) has been found. In this paper, experimental evidence is presented which indicates that ORF 4 encodes the replication protein of porcine circovirus. After cloning of the ORF 4 gene product, it was supplied in trans in a transient replication assay. The ORF 4 gene product promoted the replication of plasmid pOP11, which carries the origin of DNA replication of porcine circovirus. Since pOP11 itself is unable to replicate in virus-free porcine kidney cells, the ORF 4 gene product must be essential for replication of porcine circovirus.


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