TABLE 1.
Synthetic oligonucleotide primers
| Primer designation | Name | Sequencea | Purpose and product size (bp) |
|---|---|---|---|
| A | 3′ UL20-GFPZeo | 5′-CACGGACATCCCCCAAAACACGGGCGCCGACAACGGCAGACGATCCCTCTTGATGTTagacatgataagatacattgatgagtttgg-3′ | UL20 GET recombination (2,025) |
| B | 5′ UL20-GFPZeo | 5′-CTGACGTAAGCGACCCTTTGCGGTTTCGGTCTCCCCACCTCCACCGCACACCCCgtccgttacataacttacggtaaatgg-3′ | |
| C | 3′ gB-Kan | 5′-GTTGGGAACTTGGGTGTAGTTGGTGTTGCGGCGCTTGCGCATGACCATGTCcggttgatgagagctttgttgtaggtggac-3′ | gB GET recombination (1,209) |
| D | 5′ gB-Kan | 5′-CTGACCCTGTGGAACGAGGCCCGCAAGCTGAACCCCAACGCCATCGCCTCGTGAccacgttgtgtctcaaaatctctgatgtta-3′ | |
| E | 5′ gK-GFPZeo | 5′-AGCACCTCTCAACCGTCGTCTTGATAACGGCGTACGGCCTCGTGCTCGTGTGGTGAgtccgttacataacttacggtaaatgg-3′ | gK GET recombination (2,022) |
| F | 3′ gK-GFPZeo | 5′-CAGTTTCCAATTTGCATATGCCGTTACGGTTTCCGCCGGCCTGGATGTGACGagacatgataagatacattgatgagtttgg-3′ | |
| a | UL20-Rev | 5′-GAAGGCTGCGGCCTCGTTCCAG-3′ | Diagnostic PCR for wt UL20 (909) or ΔUL20-GFPZeo (2,469) |
| b | UL20-For | 5′-ACGGCCTTATCAAAACACTCGCCTC-3′ | |
| c | gB-Rev | 5′-TCACAGGTCGTCCTCGTCGGC-3′ | Diagnostic PCR for wt gB (2,127) or ΔgB-Kan (2,261) |
| d | gB-For | 5′-GTGATCGACAAGATCAACGCCAAGG-3′ | |
| e | gK-For | 5′-GGCACGCCATGGTCGCCGTCCGTTCCCTGC-3′ | Diagnostic PCR for wt gK (1,265) or ΔgK-GFPZeo (2,238) |
| f | gK-Rev | 5′-ATCACATACCCCGTTCCGCTTCCG-3′ |
a
For GET recombination primers, the region homologous to HSV-1 DNA is in uppercase and the region which binds to the marker gene is in lowercase letters.