TABLE 1.
Primers used for targeted deletion or disruption of S. agalactiae genes encoding putative peptide uptake systems
| Target gene | NEM316 gene designation | Type of inactivation | Primers (5′→3′)a | Cloning sites |
|---|---|---|---|---|
| oppA1 | gbs0144 | Deletion | CCGCGGATCCGTCAAGGAAGCAATAGCAGC | BamHI |
| CCCATCCACTAAACTTAAACAGCAACTGAAGCAAGTGTAAGG | ||||
| TGTTTAAGTTTAGTGGATGGGACTCTACTTCCGTAGTGG | ||||
| TGGCACAAGCTTATCATACGTGATACTGGCTG | HindIII | |||
| oppA2 | gbs0966 | Disruption | TGGCACAAGCTTTGAGTCAAAGAACTCTGG | HindIII |
| CCGCGGATCCGGACTTGTCAATTTAAAGG | BamHI | |||
| oppB | gbs0145 | Deletion | CCGCGGATCCAGATGCTGATGTTCCTGC | BamHI |
| CCCATCCACTAAACTTAAACAGCAAGAACTGGCAATTACC | ||||
| TGTTTAAGTTTAGTGGATGGGGCTATCTTGATAACAGACG | ||||
| TGGCACAAGCTTGAATACCTACGATGATGCGC | HindIII | |||
| dppA | gbs0187 | Deletion | GGGGGTACCTCGAGACAGCAATGGCGTGTATC | KpnI |
| CCCATCCACTAAACTTAAACACGCCAGAAACAAAGCAAC | ||||
| TGTTTAAGTTTAGTGGATGGGGACTTGGGATGAATCTGC | ||||
| CCGCGGATCCTCAATCAAGCAATTTCCC | BamHI | |||
| dppB | gbs0188 | Deletion | TGGCACAAGCTTTCAAGGCTAGTAACTGGG | HindIII |
| CCCATCCACTAAACTTAAACAGAGTCGAGGAAACCATTC | ||||
| TGTTTAAGTTTAGTGGATGGGGTGATACACCGCTACTTC | ||||
| CCGCGGATCCCCTTGAGCACCTTTCCCAAC | BamHI | |||
| dpsA | gbs1444 | Deletion | CCGCGGATCCCTACGAGCAAGTACACAGTC | BamHI |
| CCCATCCACTAAACTTAAACACGAAAATAGCACGCATACC | ||||
| TGTTTAAGTTTAGTGGATGGGCTCACCAGTTGGATTATCAG | ||||
| TGGCACAAGCTTCATTGGTACACAGATGG | HindIII | |||
| gbs1577 | gbs1577 | Disruption | GAGCGGGGTACCCTTACGCTTGCTTGGGGAG | KpnI |
| CCGCGGATCCCTTTTCACAACCCATTGAC | BamHI |
a
Restriction sites are underlined, and regions used for crossover PCR are in boldface type.