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.
