TABLE 3.
Plasmid design for homologous recombination
| Plasmid | Upstream flanking sequence |
Spacer | Downstream flanking sequence |
||||
|---|---|---|---|---|---|---|---|
| 5′ Restriction site (sequence and name) | BHV-1 sequence (position)a | 3′ Restriction site (sequence and name) | 5′ Restriction site (sequence and name) | BHV-1 sequence (position)a | 3′ Restriction site (sequence and name) | ||
| pBHV-1 VP22Δ | GAATTC (EcoRI) | CAAGACAAA…TCCGGCTCG (8310–9498) | TAACTGAGGTACC (Stop-Stop-KpnI)b | CCGCGC | GGATCC (BamHI) | GTTCAGCGC…GTTTTCGGG (9846–11285) | AAGCTT (HindIII) |
| pBHV-1 gMΔ | GAATTC (EcoRI) | CGGTCCGTG…GTCTCCTTA (83819–84818) | AAGCTT (HindIII) | CCGCGC | GGTACC (KpnI) | TGCCACCAG…CGCGTGACC (85188–86287) | GGATCC (BamHI) |
a
Nucleotide (nt) positions refer to GenBank accession number JX898220; VP22 open reading frame, nt 9400 to 10176; gM open reading frame, nt 84816 to 86051.
b
Stop codons are in boldface.