Nucleotide Sequencing and Expression of Type 2 Porcine Circoviruses Isolated in Korea.
- Author:
Kyung Mi KIM
1
;
Hyo Sun PARK
;
Jae Hoon KIM
;
Shien Young KANG
Author Information
1. Research Institute of Veterinary Medicine and College of Veterinary Medicine, Chungbuk National University, Cheongju, Korea. sykang@chungbuk.ac.kr
- Publication Type:Original Article
- Keywords:
Porcine circovirus type 2;
Nucleotide sequence;
ORF2;
Baculovirus expression
- MeSH:
Amino Acids;
Baculoviridae;
Base Sequence;
Blotting, Western;
Canada;
Capsid;
Circoviridae;
Circovirus*;
DNA, Circular;
Genome;
Glycosylation;
Humans;
Korea*;
Swine;
Wasting Syndrome
- From:Journal of Bacteriology and Virology
2003;33(2):151-160
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
Porcine circovirus (PCV) is a small, nonenveloped virus that contains a single-stranded circular DNA genome of about 1.76 Kb and belongs to the family Circoviridae. The PCV-2 was thought to be one of the causative agents for postweaning multisystemic wasting syndrome (PMWS) in pigs. In this study, the complete genome of two PCV-2 Korean isolates (KSY-1 and KSY-2) were sequenced and characterized. Also, the ORF2 gene of KSY-1 isolate was expressed in baculovirus expression system and the expressed protein was characterized. The sequence data indicated that the PCV-2 genome of two Korean isolates were 1,768 bases in length and encoded 2 major proteins, Rep (ORF1, 314 amino acids, 37 kDa) and a capsid (ORF2, 233 amino acids, 28~30 kDa) protein. There were 5 glycosylation sites and stem-loop structures with the nonanucleotide (5-AAGTATTAC-3), typically seen in PCV-2. Compared to nucleotide sequences of PCV-1 and PCV-2 reference strains, two Korean isolates were closely related; that is, they showed 98% homology in nucleotide sequence each other. Also, they showed 95~99% homology in nucleotide sequences with those of PCV-2 isolates but 76% similarity with those of PCV-1 reference strains. A phylogenetic analysis revealed that nucleotide sequences of Korean isolates were close to those of PCV-2 (AF055392) isolated in Canada. The baculovirusexpressed ORF2 migrated at 30 kDa and reacted with PCV-2 specific antiserum by indiect fluorescent antibody and Western blot analyses. It is concluded that our results could be valuable to understand the molecular characteristics of PCV-2 and to develop diagnostic methods for PCV-2 infections.
