Table 1.
Strains, synthetic peptides and primers used in this study.
| Strains | Descriptiona | Source or characteristics |
|---|---|---|
| Streptococcus mitis CCUG 31611T | S. mitis biovar 1 type strain, corresponds to NCTC 12261T; encapsulated, transformable strain | CCUG |
| S. mitis NCTC 8033 (SK321) | S. mitis strain isolated from the oral cavity | NCTC; (Kilian et al., 2008) |
| S. mitis CCUG 62644 (SK575) | S. mitis strain isolated from blood; ErmR | CCUG; (Kilian et al., 2008) |
| S. mitis CCUG 62641 (SK579) | S. mitis strain isolated from blood | CCUG |
| S. mitis CCUG 62642 (SK616) | S. mitis strain isolated from blood | CCUG; (Kilian et al., 2008) |
| MI009 | CCUG 31611, but ΔsrtA::erm; ErmR | This study |
| MI055 | CCUG 31611, but psigX luc::spc; SpcR | This study |
| MI074 | CCUG 31611, but ΔSM12261_0092::kan; KanR | This study |
| MI091 | CCUG 31611, but Δshp | This study |
| MI092 | SK321, but psigX luc::spc; SpcR | This study |
| Competence Stimulating Peptide (CSP) | Sequence | |
| S. mitis CCUG 31611 | EIRQTHNIFFNFFKRR | GenScript |
| S. mitis SK321 | ESRLPKIRFDFIFPRKK | GenScript |
| S. mitis SK575 | EMRKMNEKSFNFF | GenScript |
| S. mitis SK579 | EIRKSNSALVNFFK | GenScript |
| S. mitis SK616 | EMRRIDKIFINFLKRR | GenScript |
| Primers | Sequence (5′ to 3′) | |
| FP001b | ggcgcgccGTTTGATTTTTAATG | KanR cassette |
| FP015b | ggcgcgccCCGGGCCCAAAATTTGTTTGAT | ErmR cassette |
| FP016c | ggccggccAGTCGGCAGCGACTCATAGAAT | ErmR cassette |
| FP068c | aggccggccTAGGTACTAAAACAATTCATCCAGTA | KmR cassette |
| FP411 | AGCCGTTCGTGGTATGAGTC | MI009 mutant construction |
| FP412b | ggcgcgccTTTGATACCTGGCTGACTTGG | |
| FP413c | ggccggccAGATCGCGTTGATGAGATTGA | |
| FP414 | CTCAGCCGTTTTTCTCCAAG | |
| FP770d | aaagctagcAGCTGCTTTAGTCGCTGCTC | pFW5-luc |
| FP771e | aaaggatccCAATCCCCTGGACTTCTTCA | pFW5-luc |
| FP829c | ggccggccTTTTTGCTGTAAACTAAGGAGTAGAGA | MI074 mutant construction |
| FP830 | CGACTTCGATCTTTTAGTTGATG | |
| FP831 | CCCGTTTTCCCAACAAAAA | |
| FP832b | ggcgcgccTTTTCTGTACACCTCATTTCCTTT | |
| FP914 | TCTCGCGGAAGATAAAGTCG | MI009 mutant amplification |
| FP915 | GGGTTAGCAGCAACAAGGAA | |
| FP1163 | CATCTTGATAGCGTGGCTCA | |
| FP1164 | GAAGTCTAAATATGAAGAAGGTGTACACCTAAGTCT | MI091mutant construction |
| FP1165 | AGACTTAGGTGTACACCTTCTTCATATTTAGACTTC | |
| FP1166 | TTGAATTGAGACGGATTGGA | |
| FP1167 | GCTGAGTTCCTCCAAGCTGT | MI009 mutant amplification |
| FP1168 | ATCCAGAAACGACCGACAAG | |
| FP1169 | TCTTTCATCGTGTCCTCCAA | |
| FP1170 | TGATTCTCCCCGTTCAAATC | |
| FP1171 | AAACATTTTATCCACCGACAA | |
| FP1172 | TTGCACCAAGTTAGCGAATG | |
| FP1242 | CTTGAGCTGGGCTTCGTAGT | |
| FP1243 | ACAGGGGATGTCATGGGTAA | |
| FP1244 | CTGCTACGTTTGCAAGGTCA | |
| FP1245 | ATGAACGTGAACCCTTGGTC | |
| FP1248 | CAGCATGCAGTTGCAAAGTT | |
| FP1249 | CCCGAACGTGTTTCTCAAAT | |
| FP1250 | CTCAGCCGTTTTTCTCCAAG | |
| FP1251 | AGTATGGCCGTATTGCCAAG | |
| FP1280 | GCACCTGTATCACGAAGCAA | |
| FP1281 | TGCTGAAGCACAAGCTGAGT |
aKan, kanamycin; Erm, erythromycin; Spc, spectinomycin.bAscI restriction site underlined.cFseI restriction site underlined.dBamHI restriction site underlined.eNheI restriction site underlined.