TABLE 1.
Plasmids, oligonucleotides, cells, and strains used in this studya
| Name | Description | Source or reference |
|---|---|---|
| Plasmid | ||
| pCR2.1 | Kmr, Ampr | Invitrogen |
| pCB26-23 | pCR2.1 with I7L behind its native promoter | This work |
| pCB26-23-17 | pCB26-23 with aa 104 mutated Y→C and aa 324 mutated L→M | This work |
| pRB21 | puc derived with F13L flanks, MCS downstream of synthetic early-late promoter | 3 |
| pI7L | pRB21 plasmid with full-length I7L | 4 |
| pI7L-17 | pI7L with aa 104 mutated Y→C and aa 324 mutated L→M | This work |
| p78-104 | pI7L with aa 78 mutated to create an XbaI site; aa 104 mutated Y→C | This work |
| p78-324 | pI7L with aa 78 mutated to create an XbaI site and aa 324 mutated L→M | This work |
| p78-104-324 | pI7L with aa 78 mutated to create an XbaI site; aa 104 Y→C; aa 324 L→M | This work |
| Oligonucleotides | ||
| CB26 | 5′-GAG CTC GTT TTC CTA GTG ATG GAG GAG-3′ | This work |
| CB23 | 5′-AAG CTT TTA TTC ATC GTC GTC TAC-3′ | This work |
| CB84 | 5′-GAG TCG GGG CAC CTG TCT AGA CCC AAT AGT AGC G-3′ | This work |
| CB85 | 5′-CGC TAC TAT TGG GTC TAG ACA GGT GCC CCG ACT C-3′ | This work |
| CB86 | 5′-CGC GTT ACC ATT CTA TAT GTG ATG TTT TTG AGT TAC C-3′ | This work |
| CB87 | 5′-GGT AAC TCA AAA ACA TCA CAT ATA GAA TGG TAA CGC G-3′ | This work |
| CB88 | 5′-GTG GAA GTT AAT CAG CTG ATG GAA TCT GAA TGC GGG-3′ | This work |
| CB89 | 5′-CCC GCA TTC AGA TTC CAT CAG CTG ATT AAC TTC CAC-3′ | This work |
| Cell line | ||
| BSC40 | BSC1 African green monkey kidney cells adapted to grow at 40°C | 21 |
| 293 | Human embryonic kidney cells | Invitrogen |
| HeLa | Human cervical carcinoma cells | ATCC |
| L929 | Mouse fibroblast cells; ATCC CCL-1 | ATCC |
| Virus | ||
| VV WR | Western Reserve strain of vaccinia virus | ATCC |
| VV COP | Copenhagen strain of vaccinia virus | ATCC |
| VV IHDJ | IHDJ strain of vaccinia virus | ATCC |
| Cowpox | Brighton Red strain | Duke University |
| Adenovirus | Mastadenovirus, human adenovirus C | ATCC |
| MHV-A59 | Mouse hepatitis virus | ATCC |
| EMC | Encephalomyocarditis virus | ATCC |
| vvGFP | Western Reserve vaccinia virus with GFP in the TK locus | This work |
| v17-3 | vvGFP resistant to TTP-6171 with mutations in IL aa 104 and 324 | This work |
| vCB | Recombinant vvGFP with the I7L gene replaced with the mutant I7L gene | This work |
| ts16 | Temperature-sensitive VV with mutation mapped to I7L | 56 |
| Bacteria | ||
| E. coli INVαF′ | F′ endA1 recA1 hsdR17 (rK− mK+) supE44 thi-1 gyrA96rel A1O80 lacZAM15 A(lacZYA-arg-F)U16 | Invitrogen |
| P. aeruginosa | Broad spectrum of resistance to various commerical germicides | ATCC |
| S. cerevisiae | Budding yeast derived from X2180 | ATCC |
a
aa, amino acid; Tk, thymidine kinase.