| Applications |
This plasmid was used to construct CPV C62B23, ATCC VR-2209, which produces immunogenic VP-2 when inoculated into Spodoptera frugiperda cells. The BamHI linker site was used to insert the canine parvovirus VP-2 gene from strain C-780916 (CPV-916), ATCC VR-953, so that transcription of the VP-2 gene was under the control of the polyhedrin gene promoter. |
| Comments |
Restriction digests of the clone give the following sizes (kb): HindIII--6.7, 3.7, 1.1; BamHI--9.1, 2.2; PvuII--3.7, 3.6, 2.2, 1.6, 0.6; EcoRI/EcoRV--3.7, 3.7, 2.5, 1.4. The insert is from nt 2750 to nt 4990 of the GenBank sequence, but it differs from the GenBank sequence at the following nt (GenBank to pAc6C2B23 change): 3065 (from C to A), 3094 (from T to A), 3480 (from A to G), 3855 (from C to T), 3596 (from T to C), 3657 (from C to T), 3753 (from A to G). This plasmid was used to construct CPV C62B23, ATCC VR-2209, which produces immunogenic VP-2 when inoculated into Spodoptera frugiperda cells. A 7.0 kb EcoRI fragment from AcNPV 1A strain, containing the polyhedrin gene, was inserted into the EcoRI site. Nucleotides -7 to +461 were deleted (where +1 is the A of the polyhedrin ATG), and a 23 base BamHI linker was inserted. This vector provides the strong polyhedrin promoter, polyhedrin terminator signals, and sufficient flanking sequence to promote homologous recombination in cells co-transfected with baculovirus. Constructed from a pUC19 derivative from which the BamHI recognition site had been removed. The BamHI linker site was used to insert the canine parvovirus VP-2 gene from strain C-780916 (CPV-916), ATCC VR-953, so that transcription of the VP-2 gene was under the control of the polyhedrin gene promoter. |
| References |
Wood HA, Parrish CR. Subunit canine parvovirus vaccine. US Patent 4,971,793 dated Nov 20 1990
Luckow VA, Summers MD. Trends in the development of baculovirus expression vectors. Bio-Technology 6: 47-55, 1988.
Smith GE, et al. Production of human beta interferon in insect cells transfected with a baculovirus expression vector. Mol. Cell. Biol. 3: 2156-2165, 1983. PubMed: 6318086
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