Recombination virus particles for expressing 2-typed porcine circovirus nucleocapsid protein Cap gene
A nucleocapsid protein, porcine circovirus technology, applied in the direction of antiviral agents, genetic engineering, virus antigen components, etc., can solve the problems of difficult to cure PMWS, unclear mechanism, and no clinical symptoms of the virus, and achieve replication. The effect of high efficiency and large clone space
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[0044] 1. Construction and application of PPV VP2 virus-like particles expressing foreign genes
[0045] 1) Obtain the ΔVP2 target gene and clone it into pMD19-T vector
[0046] Using the NADL-2 gene sequence in GenBank with accession number NC: 001718 as a template, primers were designed with the software Primer 5.0: KpnI and NotI restriction sites were added, respectively. Attenuated PPV strain (NADL-2) was purchased from China Veterinary Drug Control Institute.
[0047] P1 5′-gaggtaccgggggggttggtgtgt-3′
[0048] P2 5′-cgggcggccgcctagtataattttcttgg-3′
[0049] With P1, P2 primer amplifies the fragment containing VP2 (1740bp) gene C-terminal 1640bp ( figure 1 ). After the amplified PCR product was electrophoresed on 1% agarose gel, it was recovered and connected with pMD19-T Vector to transform E.coli DH 5 α, Plasmid was extracted by alkaline lysis method, identified by KpnI and Not I double enzyme digestion, and a positive plasmid (pTΔVP2) was obtained.
[0050] 2) Con...
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