title: Instability in vitro of a PCV2 infectious clone containing an insertion between ORF1 and ORF2.
abstract: Porcine circovirus 2 (PCV2) is a small circular single-stranded DNA virus that causes postweaning multisystemic wasting syndrome in pigs. We obtained an infectious clone, derived from PCV2 strain 1147 (size 1,767 bp), with a 60 bp insertion in the cap-rep intergenic region at the 3' end of the rep. In vitro propagation and sequencing of virus showed mixed sequence for the cap-rep intergenic region. Cloning of this region of virus from the passages 2 and 20 showed deletions of various size of the 60 bp insertion. Viruses from the passage 20 showed deletions of the 60 bp insertion and had extra (+1 to +5) or less (-1 or -2) bp in the intergenic region as compared to 1,147. These findings suggest that insertion of a 60-bp DNA sequence at the 3' end of the rep is unstable in vitro. This finding can have implications for the genetic engineering of PCV2.
authors: Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium. Dipongkor.saha@ugent.be
authors: Dipongkor
authors: D
authors: Saha
authors: Jan V
authors: JV
authors: Doorsselaere
authors: Hans J
authors: HJ
authors: Nauwynck
mesh_terms: D000818:Animals; D017925:Circovirus / Q000235:genetics* / Q000254:growth & development*; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods; D042822:Genomic Instability; D016254:Mutagenesis, Insertional; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D012692:Serial Passage; D013552:Swine; D014773:Virology / Q000379:methods
chemical_list: D004279:DNA, Viral