The PCV-2 positive tissues by PCR-restriction fragment length polymorphism (PCR-RFLP) were inoculated to the PK-15 cells not contaminated by PCV. The viral particles were observed in the infected PK-15 cells by eletro...The PCV-2 positive tissues by PCR-restriction fragment length polymorphism (PCR-RFLP) were inoculated to the PK-15 cells not contaminated by PCV. The viral particles were observed in the infected PK-15 cells by eletronmicroscope .It was concluded that the isolate was PCV-2 and was named SCH-A strain.The capsid protein gene(cp) in ORF2 was amplified from PK-15 cell line infected with SCH-A strain by PCR, and was cloned and sequenced.The nucleotide sequences of ORF2 gene of the SCH-A strain was compared with other porcine circovirus isolates.The results showed that the homologies of nucleotides were between 90.5% and 96.7% in comparision with the PCV-2 reference strains,but the homologies of nucleotides were only about 67% with the PCV-1 reference strains. SCH-A strain displayed the highest nucleotide homology((96.7)%) with the reference strain Hunan .Phylogenetic tree analysis showed ORF2 gene of the SCH-A isolate belongs to PCV-2 .展开更多
PCV1 was isolated from IBRS-2 cells line,and its complete genomic sequence was cloned by PCR.Sequence analysis indicated that this PCV1 strain shares >98% nucleotide identity with the other PCV1 strains in GenBank....PCV1 was isolated from IBRS-2 cells line,and its complete genomic sequence was cloned by PCR.Sequence analysis indicated that this PCV1 strain shares >98% nucleotide identity with the other PCV1 strains in GenBank.Then double copy molecule clone(pSK2PCV1) was constructed and used to transfect PK-15 cell line.The results of indirect immunofluorescence(IIF) and RT-PCR suggested that pSK2PCV1 could form infectious virus after being transfected into PK-15 cells.展开更多
文摘The PCV-2 positive tissues by PCR-restriction fragment length polymorphism (PCR-RFLP) were inoculated to the PK-15 cells not contaminated by PCV. The viral particles were observed in the infected PK-15 cells by eletronmicroscope .It was concluded that the isolate was PCV-2 and was named SCH-A strain.The capsid protein gene(cp) in ORF2 was amplified from PK-15 cell line infected with SCH-A strain by PCR, and was cloned and sequenced.The nucleotide sequences of ORF2 gene of the SCH-A strain was compared with other porcine circovirus isolates.The results showed that the homologies of nucleotides were between 90.5% and 96.7% in comparision with the PCV-2 reference strains,but the homologies of nucleotides were only about 67% with the PCV-1 reference strains. SCH-A strain displayed the highest nucleotide homology((96.7)%) with the reference strain Hunan .Phylogenetic tree analysis showed ORF2 gene of the SCH-A isolate belongs to PCV-2 .
文摘PCV1 was isolated from IBRS-2 cells line,and its complete genomic sequence was cloned by PCR.Sequence analysis indicated that this PCV1 strain shares >98% nucleotide identity with the other PCV1 strains in GenBank.Then double copy molecule clone(pSK2PCV1) was constructed and used to transfect PK-15 cell line.The results of indirect immunofluorescence(IIF) and RT-PCR suggested that pSK2PCV1 could form infectious virus after being transfected into PK-15 cells.
