TABLE 2.
Primers used for generation of recombinant HSV-BACsa
| BAC | Mutation | Primer sequence |
|---|---|---|
| KOS BAC | UL6 ORF deletion + insertion of RNA Pol II promoter for UL5 gene | 5’ CTGTCCGTCTAGCTGGCGCTCCCCGCCGGCCGCCGCCATGGCTGGCGGCGGACGCTTCGGTAGTGAGCTTGTCGGTCCCTTAGCGCCTTTATATTGGGTGCCTAGCCGGTCGACCGCCCTTAGGGATAACAGGGTAATCGATTT 3’ |
| 5’ GGCGATGGCCTGCTTGAGGATGGTGGCGGCCGACCCCTCCATAGGGCGGTCGACCGGCTAGGCACCCAATATAAAGGCGCTAAGGGACCGACAAGCTCACTACCGAAGCGTCCGCCGCCAGCCGCCAGTGGTACAACCAATTAACC 3’ | ||
| KOS BAC | UL17 ORF deletion | 5’ CCGCATCGCCTACACACCCACACCCACCCCCGAACCCGGGCGCCTTCCCCCGGCTAGGGATAACAGGGTAATCGATTT 3’ |
| 5’ GAGTGGGTGGGCGAGGTGGCCGGGGGAAGGCGCCCGGGTTCGGGGGTGGGTGTGGCCAGTGTTACAACCAATTAACCA 3’ | ||
| KOS BAC | UL36 Cterm-GFP | 5’ CGTGCTGACCAGCCTACATCACGTGCGCATGTTACTGGGCGTGAGCAAGGGCGAGGAGCTG 3’ |
| 5’ CGGTTCGAAACTTAACAACAAAATAATCGAGCGCGTCTACTTGTACAGCTCGTCCATGCCG 3’ | ||
| bFH422 | UL25-GFP and UL36-RFP | 5’ CGTGCTGACCAGCCTACATCACGTGCGCATGTTACTGGGCGCCTCCTCCGAGGACGTCATC 3’ |
| 5’ CGGTTCGAAACTTAACACACAAAATAATCGAGCGCGTCTACAAGGCGCCGGTGGAGTGGCG 3’ | ||
| KOS BAC | UL17i | 5’ GACGAGCTCGTCCCCCCCAACACGCGCTACGCGTGCGGCCGCAACGCGGCCGACAGCACGCGCATCTAGGGATAACAGGGTAATCGATTT 3’ |
| 5’CCGACAGACGCGCATGATGCGCGTGCTGTCGGCCGCGTTGCGGCCGCACGCGTAGCGCGTGTTGGGGCCAGTGTTACAACCAATTAACCA 3’ |
a
Abbreviations and symbols: Pol, polymerase; Cterm, C terminal; underlining, HSV sequence; italic, Pol II promoter sequences; bold, template sequence (pEP-Kan-S, pEP-GFP-in, pEP-RFP-in).