TABLE 1.
Primers used in this study
| Primer | Sequencea | Descriptionb |
|---|---|---|
| BFW050 | CCGGAATTCCGGCGAAATTAGAAAAGAGGACAAGCATATGAAAAAAGGTTTTTTTCTCATGGCTATGGTCGTGAGT | Reverse amplification primer for lsp |
| BFW052 | CCGCTGCAGTTACTTCAACTGTTGATAGAGCACTTCCAAATTTGCTCTAAGATTTTCTAGATATGTCTTGTTTCCG | Forward amplification primer for lsp |
| BFW053 | GTGAGTTTAGTAATGATAGCAGGGGCTGATAAGTCAGCAAACCCCAAACAG | Reverse deletion primer for lspC20A |
| BFW054 | CTGTTTGGGGTTTGCTGACTTATCAGCCCCTGCTATCATTACTAAACTCAC | Forward deletion primer for lspC20A |
| BFW055 | GATGATCCAATCAGGTGCAGGCATTGCTTCCTTTGAACCGTCTGTAAATGATG | Reverse deletion primer for lspH66A |
| BFW056 | CATCATTTACAGACGGTTCAAACCAAGCAATGCCTGCACCTGATTGGATCATC | Forward deletion primer for lspH66A |
| BFW057 | TGATCCTGCGACACTTTATGACCCAGCTACCTGGACAGATCCCGTTTTAGCTG | Reverse deletion primer for lspH142A |
| BFW058 | CAGCTAAAACGGGATCTGTCCAGGTAGCTGGGTCATAAAGTGTCGCAGGATCA | Forward deletion primer for lspC20A |
| BFW068 | GCGCGGTACCAACTAAACCAGAAAAAGATAGTTC | Reverse amplification primer paired with BFW052 for pJRS233 cloning of lspH66A |
| BFW069 | GCGCGGTACCAGTTGCCATTGACAAAGCATTGAT | Reverse amplification primer paired with BFW052 for pJRS233 cloning of lspH142A |
a
Restriction sites engineered for cloning purposes are underlined.
b
Primers are described according to their purposes as follows: “amplification” primers amplify the region containing the gene of interest for construction of mutant S. pyogenes strains, and “deletion” primers were used to amplify genes of interest for cloning into a streptococcal expression vector as described in Materials and Methods.