TABLE 2.
Primers used in the construction and testing of live vector strains and expression plasmids
| Primer | Sequencea | Template | Reference |
|---|---|---|---|
| 1 | 5′-CATATGATTGACCTGAATGAATATACAGTATTGGAA-3′ | ssb locus from S. flexneri 2a strain 2457T | 42 |
| 2 | 5′-GCTAGCTATTGTTTTAATGACAAATCAGAACGGAA-3′ | ssb locus from S. flexneri 2a strain 2457T | 42 |
| 3 | 5′-CTCGAGCTTGCCAGATTTTCCAGCGTTTTGGTGTGT-3′ | Ty2 | 8 |
| 4 | 5′-CATATGTTATTATTATTAGCTAGCTACTGTATATTCAAACAGGTTAAATTGTGT-3′ | Ty2 | 8 |
| 5 | 5′-CATATGCATTTTCGCTATAGTTCTCGTCTGCTGAAA-3′ | Ty2 | 8 |
| 6 | 5′-CTCGAGACTAGTTAGCTAATCATTGAAACTCTAAATCATTTT-3′ | Ty2 | 8 |
| 7 | 5′-CATATGAATATCCTCCTTAGTTCCTATTCC-3′ | pKD3 | 7 |
| 8 | 5′-GCTAGCGTGTAGGCTGGAGCTGCTTCGAAGTTCTA-3′ | pKD3 | 7 |
| 9 | 5′-TTCGGCGGATCGGAGAGATCGCAGACTTCG-3′ | Ty2 | 8 |
| 10 | 5′-AGACATCAATTATTGCACTAACTATATCTT-3′ | Ty2 | 8 |
| FRT-tetA-for | 5′-TCTAGAgaagttcctattctatatatagtataggaacttcGCTAGCTCATGTTTGAC AGCTTATCATCGATAAGCTTTAATGCGGTAGTTTATCAC-3′ | pSEC91 | 13 |
| FRT-tetA-rev | 5′-TCTAGAgaagttcctatactatatatagaataggaacttcGCTAGCCTATCAGGTCGAGGTG GCCCGGCTCCATGCACCGCGACGCAACGCGGGGAG-3′ | pSEC91 | 13 |
| gfpuv-for | 5′-CAGTGGAGAGGGTGAAGGTGATGC-3′ | pGEN222S2 | This work |
| gfpuv-rev | 5′-GTGTCGACAGGTAATGGTTGTCTG-3′ | pGEN222S2 | This work |
| ssb-for | 5′-GTCAGGACCCGGAAGTACGCTACATGCCAAATGGTGGCGCAG-3′ | pGEN222S2 | This work |
| ssb-rev | 5′-CGCCGCTGAACTGATTGCCACCCTGCGGCTGCTGAGGCTGAC-3′ | pGEN222S2 | This work |
| ori15A-for | 5′-GTCCTTCTTGAGTTTGTAACTGCTGCTG-3′ | pGEN222S2 | This work |
| ori15A-rev | 5′-CGAGGTAACTGGCTTGGAGGAGCGCAGTCAC-3′ | pGEN222S2 | This work |
a
Relevant restriction sites are underlined. The FRT recombination sites flanking the tetA gene are in lowercase.