TABLE 1.
Plasmids and primers used in the study
| Plasmid or primer | Descriptiona | Reference, source, or comment |
|---|---|---|
| Plasmids | ||
| pRAD1 | E. coli-D. radiodurans shuttle vector (copy no., 7-10 per cell in D. radiodurans) | 28 |
| pRN1 | pRAD1 with phoN cloned at NdeI and BamHI sites; Cbr Cmr | Lab collection |
| pTZ57R/T | PCR product cloning vector (2,886 bp, Apr) for blue-white selection of transformants | MBI Fermentas |
| pTAPssb102 | 102-bp sequence upstream of D. radiodurans ssb cloned in pTZ57R/T | This study |
| pSN2 | pRN1 with 112-bp fragment from pTAPssb102 subcloned at NruI and NdeI sites | This study |
| pSN3 | pRN1 with 132-bp sequence upstream of D. radiodurans ssb ORF cloned at XbaI and NdeI sites | This study |
| pSN4 | pRN1 with 351-bp sequence upstream of D. radiodurans ssb ORF cloned at XbaI and NdeI sites | This study |
| Primers | ||
| P1 | 5′-CCGAGAAGGATTACAATCTAGAACG-3′ | XbaI |
| P2 | 5′-CGGAGTGGAAGATCTAGAAGGCCTG-3′ | XbaI |
| P3 | 5′-GCGAGCTCATAATTGACTCTGCTTGTTACTATCTAGTG-3′ | SacI |
| P4 | 5′-CATGCCTCGGGCCATATGAAATTCT-3′ | NdeI |
| P5 | 5′-GGAGCGGATAACAATTTCACACA-3′ | P5 and P6 flank multiple-cloning site, 58 bp on either side in pRad1 |
| P6 | 5′-AACGCGGCTGCAAGAATGGTA-3′ |
a
The underlined sequence corresponds to the restriction site incorporated in the primer.