TABLE 9.
Oligonucleotide sequences and DNA templates for the construction of plasmids
| Constructed plasmid | Oligonucleotide sequence (5′–3′) | PCR DNA template (reference or source) | Recipient plasmid (source) |
|---|---|---|---|
| pBS-UL7 | GATAAGCTTGATATCGAATTCACACCGCGTCCGTCAAGGTGC | pYEbac861Cre (this study) | pBluescript II KS(+) (Stratagene) |
| CGCTCTAGAACTAGTGGATCGACGGCGCGGTTTTGTTTCTC | |||
| pBS-UL51 | GCGAATTCGTCCGTTCTGTCGTCGCTAT | Wild-type HSV-2 186 genome | pBluescript II KS(+) (Stratagene) |
| GCAAGCTTGGTACGCGTCGTAGCTGAAT | |||
| pRetroX-TRE3G-UL7 | GCGCGGCCGCGCCACCATGGCCGCCGCGACGGCCGAC | Wild-type HSV-2 186 genome | pRetroX-TRE3G (Clontech) |
| GCGCGGCCGCTTAGCAAAACCGATAGAAAAGC | |||
| pAcGHLT-HSV-2-Us3 | GCGCGGCCGCTCACTTAGGGTGGAATAGCG | YK813 (HSV-2 Us3-repair) genome (22) | pAcGHLT-B (Pharmingen) |
| GCGAATTCTGGCCTGTCGTAAGTTCTGTGG | |||
| pAcGHLT-HSV-2-Us3-K220M | GCGCGGCCGCTCACTTAGGGTGGAATAGCG | YK811 (HSV-2 Us3-K220M) genome (22) | pAcGHLT-B (Pharmingen) |
| GCGAATTCTGGCCTGTCGTAAGTTCTGTGG | |||
| pFlag-HSV-1-Us3 | GCGAATTCAGCCTGTCGTAAGTTTTGTCG | pBS-Us3 (60) | pFlag-CMV-2 (Sigma-Aldrich) |
| GCGATATCTCATTTCTGTTGAAACAGCG | |||
| pFlag-HSV-1-Us3-K220M | GCGAATTCAGCCTGTCGTAAGTTTTGTCG | pBS-Us3-K220M (60) | pFlag-CMV-2 (Sigma-Aldrich) |
| GCGATATCTCATTTCTGTTGAAACAGCG | |||
| pFlag-HSV-2-Us3 | GCGAATTCGGCCTGTCGTAAGTTCTGTGG | pBS-HSV-2-Us3 (58) | pFlag-CMV-2 (Sigma-Aldrich) |
| GCGATATCTCACTTAGGGTGGAATAGCG | |||
| pFlag-HSV-2-Us3-K220M | GCGAATTCAGCCTGTCGTAAGTTCTGTGG | pAcGHLT-HSV-2-Us3-K220M (this study) | pFlag-CMV-2 (Sigma-Aldrich) |
| GCGATATCTCACTTAGGGTGGAATAGCG | |||
| pFlag-ChHV-Us3o-K220M | |||
| 1st PCR-A | GCAAGCTTGCATGTAGGAAGTT | pUC57-ChHV-Us3o (this study) | pFlag-CMV-2 (Sigma-Aldrich) |
| TGCTGGCATACCAGCCGGCCATCACGATCACTCTCTGTGGG | |||
| 1st PCR-B | CCCACAGAGAGTGATCGTGATGGCCGGCTGGTATGCCAGCA | ||
| GCGAATTCTCACTTTGGATGAA | |||
| 2nd PCR | GCAAGCTTGCATGTAGGAAGTT | Mixture of 1st PCR fragments | |
| GCGAATTCTCACTTTGGATGAA | |||
| pMAL-HSV-2-UL7 | GCGTCGACATGGCCGACCCCACGCCCGC | pYEbac356 (22) | pMAL-c (New England BioLabs) |
| GCAAGCTTTTAGCAAAACCGATAGAAAAG | |||
| pMAL-HSV-2-UL34 | GCGAATTCATGGCGGGGATGGGGAAGCCCTAC | pYEbac356 (22) | pMAL-c (New England BioLabs) |
| GCGTCGACTCATAGGCGCGCGCCAACCGCC | |||
| pcDNA-Myc-HSV-2-UL34 | GCGAATTCTGCGGGGATGGGGAAGCCCTA | pMAL-HSV-2-UL34 (this study) | pcDNA-Myc (this study) |
| GCGTCGACTCATAGGCGCGCGCCAACCGCC | |||
| pcDNA-SE-HSV-2-UL7 | GCGATATCAAATGGCCGACCCCACGCCCGCCG | pMAL-HSV-2-UL7 (this study) | pcDNA-SE (this study) |
| GCCTCGAGTTAGCAAAACCGATAGAAAAG | |||
| pcDNA-Myc-HSV-2-UL7 | GCGATATCGCCACCATGGAGCAGAAGCTGATCTCAGAGGAGGACCTGGCCGACCCCACGCCCGCCGA | pYEbac356 (22) | pcDNA3.1(+) (Thermo Fisher Scientific) |
| GCCTCGAGTTAGCAAAACCGATAGAAAAG | |||
| pcDNA-SE-ChHV-UL7o | GCGAATTCGCCGACCCAACCCCCGCAGA | pcDNA-Myc-ChHV-UL7o (this study) | pcDNA-SE (this study) |
| GCGATATCTTAACAAAACTGGTAAAACAGG | |||
| pEGFP-HSV-1-gB770–904 | GCGAATTCTTCATGTCCAACCCCTTTGG | Wild-type HSV-1(F) genome | pEGFP-C2 (Clontech) |
| GCGGATCCTCACAKUGGTCGTCCTCGTCGG | |||
| pcDNA-Myc-HSV-2-UL7-T288A | GCGATATCGCCACCATGGAGCAGAAGCTGATCTCAGAGGAGGACCTGGCCGACCCCACGCCCGCCGA | pYEbac356 (22) | pcDNA3.1(+) (Thermo Fisher Scientific) |
| GCCTCGAGTTAGCAAAACCGATAGAAAAGCGACGAGGCCCGTCG | |||
| pcDNA-Myc-HSV-2-UL7-S289A | GCGATATCGCCACCATGGAGCAGAAGCTGATCTCAGAGGAGGACCTGGCCGACCCCACGCCCGCCGA | pYEbac356 (22) | pcDNA3.1(+) (Thermo Fisher Scientific) |
| GCCTCGAGTTAGCAAAACCGATAGAAAAGCGACGCGGTCCG | |||
| pcDNA-Myc-HSV-2-UL7-S290A | GCGATATCGCCACCATGGAGCAGAAGCTGATCTCAGAGGAGGACCTGGCCGACCCCACGCCCGCCGA | pYEbac356 (22) | pcDNA3.1(+) (Thermo Fisher Scientific) |
| GCCTCGAGTTAGCAAAACCGATAGAAAAGCGCCGAGGTCCGTCG | |||
| pcDNA-Myc-HSV-2-UL7-SS/AA | GCGATATCGCCACCATGGAGCAGAAGCTGATCTCAGAGGAGGACCTGGCCGACCCCACGCCCGCCGA | pYEbac356 (22) | pcDNA3.1(+) (Thermo Fisher Scientific) |
| GCCTCGAGTTAGCAAAACCGATAGAAAAGCGCCGCGGTCCGTCG | |||
| pcDNA-Myc-HSV-2-UL7-TSS/AAA | GCGATATCGCCACCATGGAGCAGAAGCTGATCTCAGAGGAGGACCTGGCCGACCCCACGCCCGCCGA | pYEbac356 (22) | pcDNA3.1(+) (Thermo Fisher Scientific) |
| GCCTCGAGTTAGCAAAACCGATAGAAAAGCGCCGCGGCCCGTCG | |||
| pcDNA-SE-ChHV-UL7o-SS/AA | GCGAATTCGCCGACCCAACCCCCGCAGA | pcDNA-Myc-ChHV-UL7o (this study) | pcDNA-SE (This study) |
| GCGATATCTTAACAAAACTGGTAAAACAGGGCTGCTGTCCGCCGCTTCCTATTGC | |||
| pEGFP-AncA-UL7o-Q286R | ATGCAAGCTTGAATTCGCCG | pEGFP-AncA-UL7o (this study) | pEGFP-C2 (Clontech) |
| GCCTCGAGTCAACAAAACTGATAGAACAGGCTAGATGTTCGGCGCTTAGGGGTCTCGGAC | |||
| pEGFP-AncA-UL7o-QSS/RAA | ATGCAAGCTTGAATTCGCCG | pEGFP-AncA-UL7o (this study) | pEGFP-C2 (Clontech) |
| GCCTCGAGTCAACAAAACTGATAGAACAGGGCAGCTGTTCGGCGCTTAGGGGTCTCGGAC | |||
| pEGFP-AncB-UL7o-SS/AA | ATGCAAGCTTGAATTCGCCG | pEGFP-AncB-UL7o (this study) | pEGFP-C2 (Clontech) |
| GCCTCGAGTCAGCAAAACTGATAGAACAGAGCGGCTGTGCGCCTCTTAGGATTGC | |||
| pEGFP-HSV-1-gB770–904-T887A | |||
| 1st PCR-A | GCGAATTCTTCATGTCCAACCCCTTTGG | Wild-type HSV-1(F) genome | pEGFP-C2 (Clontech) |
| GGGAACTTGGGTGTAGTTGGCGTTGCGGCGCTTGCGCATGA | |||
| 1st PCR-B | TCATGCGCAAGCGCCGCAACGCCAACTACACCCAAGTTCCC | ||
| GCGGATCCTCACAGGTCGTCCTCGTCGG | |||
| 2nd PCR | GCGAATTCTTCATGTCCAACCCCTTTGG | Mixture of 1st PCR fragments | |
| GCGGATCCTCACAGGTCGTCCTCGTCGG |