TABLE 1.
Oligonucleotide primers
| Primer | Sequence (5′ to 3′)a | 5′ startb | 3′ endb | Use |
|---|---|---|---|---|
| 21pe1 | CTTCTTGTACTTTCAAGTTAC | 30808 | 30828 | Primer extension |
| 21pe2 | GTTTTCTCTGGTGACCATGG | 30853 | 30862 | Primer extension |
| 21pe3 | GGAATTTTCTGGGTAGATCG | 30897 | 30916 | Primer extension |
| 21-mlu | GCGACGCGTAGCTGAGGGGTTAAATTCACA | 30475 | 30497 | 5′-end p21, p21Δ |
| 21-mluA | GCGACGCGTGGTAGGAGGAGCC | 30585 | 30597 | 5′-end p21A |
| 21-mluB | GCGACGCGTGGTTCTTCAACTTACCGTG | 30626 | 30644 | 5′-end p21B |
| 21-mluC | GCGACGCGTGCGTTTTTATTGATGTTAC | 30663 | 30682 | 5′-end p21C |
| 21-mluZ | GCGACGCGTCGGGGTCACGTCCAGCCTGTG | 29979 | 30000 | 5′-end p21Z |
| 21-kpn | GCGGGTACCGGTATATTCTACGCTGACTTAAC | 30758 | 30736 | 3′-end p21, p21A, p21B, p21C |
| 21-kpnA | GCGGGTACCGGCTCCTCCTACC | 30597 | 30585 | 3′-end p21Δ |
| KpnI-MluI-p1 | TACGCGTGGAATTCCGGTACCG | NAc | NA | Modify pCAT3basic |
| KpnI-MluI-p2 | CGCGCGGTACCGGAATTCCACGCGTAGTAC | NA | NA | Modify pCAT3basic |
| 4-HindIII | CAAAATATCTGACAA(GC)TTGCGTGTTTGCAG | 4147 | 4175 | Introduce HindIII site 5′ of ORF 4 |
| 4-SpeI | CAAATTAGTATGTTTTGAC(TAGT)AGCATGAAAAAGG | 2739 | 2771 | Introduce SpeI site 3′ of ORF 4 |
| 10-EcoRI | CTTATTTAAACTAAAGA(A)TT(C)TTACTCTATAAG | 12128 | 12158 | Introduce EcoRI site 5′ of ORF 10 |
| 10-XbaI | CGCGTTAAACGTC(TAG)ATTGGGGTAGAG | 13385 | 13409 | Introduce XbaI site 3′ of ORF 10 |
| 61-HindIII | GAATACAGCCAA(G)CTTGTTACCATGG | 104505 | 104482 | Introduce HindIII site 5′ of ORF 61 |
| 61-SpeI | GAAGTCCTAGTT(AC)T(A)GTTGGGAGGGGG | 103091 | 103067 | Introduce SpeI site 3′ of ORF 61 |
| 62-EcoRI | GGGTACGTCTA(G)AATTCACCCCAG | 109160 | 109138 | Introduce EcoRI site 5′ of ORF 62d |
| 63-HindIII | GGTGCAAAACATGTCC(AAGC)TTGGGGCCGTAGTA | 110592 | 110563 | Introduce HindIII site 5′ of ORF 63 |
| 63-SpeI | TGTATTTATTTATAA(CT)AG(T)ACTACACGCCATGGG | 111435 | 111404 | Introduce SpeI site 3′ of ORF 63 |
| EcoRI-HindIII-p1 | AATTCCTGGA | NA | NA | Link EcoRI site to HindIII site in pCIneo |
| EcoRI-HindIII-p2 | AGCTTCCAGG | NA | NA | Link EcoRI site to HindIII site in pCIneo |
MluI sites are indicated by a single underline. KpnI sites are indicated by a double underline. Parentheses enclose nucleotides added to introduce the desired restriction endonuclease.
Nucleotide position of the oligonucleotide on the VZV genome.
NA, not available.
The 3′ cloning site (XbaI) was supplied by the pAlter-1 vector.