TY - JOUR
T1 - Instability in vitro of a PCV2 infectious clone containing an insertion between ORF1 and ORF2
AU - Saha, Dipongkor
AU - Doorsselaere, Jan V.
AU - Nauwynck, Hans J.
PY - 2012/4/1
Y1 - 2012/4/1
N2 - 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. © Springer Science+Business Media, LLC 2011.
AB - 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. © Springer Science+Business Media, LLC 2011.
KW - Infectious clone
KW - Porcine circovirus type 2 (PCV2)
KW - Single-stranded DNA
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U2 - 10.1007/s11262-011-0693-6
DO - 10.1007/s11262-011-0693-6
M3 - Article
C2 - 22116682
SN - 0920-8569
VL - 44
SP - 258
EP - 261
JO - Virus Genes
JF - Virus Genes
IS - 2
ER -