TABLE 4.
PCR primers for generation of BHV-1 UL49.5 C42S/VP22Δ mutant
| Primer function and name (restriction site) | Sequencea | Position (nt)b |
|---|---|---|
| Cloning | ||
| VP22/34-149 del F (EcoRI) | 5′-GCGAATTCCGCAAGACAAAGCGGCAGGGCTCC-3′ | 8308–8331 |
| VP22/34-149 del R (KpnI) | 5′-GCGCTGGGTACCTCAGTTACGAGCCGGACTCGTAGTCATAGAGGCTG-3′ | 9471–9498 |
| Verificationc | ||
| UL49.5 F | 5′-AGCGAGTCGGGCTCACAGCAGC-3′ | 8917–8938 |
| UL49.5 R | 5′-AACCGGGCCATGGCAAGCGAGTC-3′ | 9388–9410 |
a
BHV-1-specific sequences are shown in boldface letters, italicized sequences are integrated restriction sites, and underlined sequences are integrated stop codons.
b
Positions are based on GenBank accession number JX898220.
c
Primers used for verification of the recombinant BHV-1 by sequencing.