Studies On The Identification Of Recombinant Virus VFV282 With Nucleic Acid Probe And PCR Methods

The recombinant Fowlpox virus vFV282 was identified by probe hybridization and PCR.By EcoRV and Xba I digestion, the plasmid which contains NDV F gene was cut into a cDNA fragment of 344bp. Then the fragment was labeled by digoxigenin. It was described that the probe could hybridize specifically with the plasmids containing NDV F gene, but no hybridization signal was detected in other specimens such as control plasmid pUTA-2 , FPV 282E4 and CEF DNA . The sensitivity detection indicated that the probe could detect a minimal 4pg homology DNA. The probe could strongly hybridize with the recombinant Fowlpox virus vFV282. The result indicated that the probe could be used in screening and identifying of the recombinant Fowlpox virus vFV282.According to the NDV F gene and IBDV VP0 gene, we designed and composed two pairs of primers (P1, P2 and P3,P4). The specificity assay indicated that P1 and P2 were specific to the recombinant plasmid which contained NDV F gene and P3 and P4 were specific to the recombinant plasmid which contained EBDV VPO gene. But the control plasmid pUTA-2 and FPV 282E4 were all negative. The sensitivity assay indicated that the P2 could detect at least 13pg pEX IBDWPO DNA. The recombinant Fowlpox virus vFV282 could be respectively amplified by P1 and P2, P3 and P4. The result indicated that the primers P1 and P2, P3 and P4 could be used in screening and identifying the recombinant Fowlpox virus vFV282.Postgraduate: Jia Leili Major: Preventive veterinary science Advisor: Prof. Wei Ping, Prof. Jin Ningyi...