Table 1.
Bacterial strains, plasmids and primers used in this study
| Strains/plasmid | Relevant characteristics | References |
|---|---|---|
| S. mutansUA159 | Wild-type | (Ajdic et al., 2002) |
| S. mutans TW26D | ΔluxS, Kanr | This study |
| S. mutans TW297.1 | Δsmu.44, Kanr | This study |
| S. mutans TW238 | Δsmu.423, Kanr | This study |
| S. mutans TW294.1 | Δsmu.46, Kanr | This study |
| S. mutans TW310 | Δsmu.299, Kanr | This study |
| pDL278 | Shuttle vector, Spr | (LeBanc & Lee, 1991) |
| pDL278:luxS | Shuttle carrying luxS, Spr | This study |
| S. mutans TW26DC | TW26D/pDL278:luxS, Ermr and Spr | This study |
| pAYBG754S | Shuttle vector with a gfp reporter, Kanr | (Yoshida & Kuramitsu, 2002) |
| Primers for mutagenesis | Sequence (5′–3′) | Sequence (5′–3′) | Flanking region amplified |
|---|---|---|---|
| luxS 5F | 55-aaatctgtcattgctgatggac | 53-atacccgagctcctactgagtagtc | 5′ of luxS for mutagenesis |
| luxS 3F | 35-agttcaaagctttcaacagtaacttc | 33-tgctagacgttgcatagcttgagc | 3′ of luxS for mutagenesis |
| smu.44 5F | 55-tgttggaaatccaccatatattacgtatc | 53-tgcacttctgaattctagtacattc | 5′ of smu.44 for mutagenesis |
| smu.44 3F | 35-ttcataacaggagaattcttagatac | 33-tccttcttcactaaagtgttgtac | 3′ of smu.44 for mutagenesis |
| smu.46 5F | 55-agatagtcttgttaacaaacatc | 53-agtttgccagaattcaatagcatc | 5′ of smu.46 for mutagenesis |
| smu.46 3F | 35-tagaacaccagaattctattaatcaac | 33-acgaccaccagaaccaacaac | 3′ of smu.46 for mutagenesis |
| smu.423 5F | 55-tcactattactgatgtccaaatg | 53-ttgttcaaatgcatgcgtattcat | 5′ of smu.423 for mutagenesis |
| smu.423 3F | 35-tgtaggtgcatgcactttttgttg | 33-ttcttcactcattttgatgtc | 3′ of smu.423 for mutagenesis |
| smu.299 5F | 55-atgattcatctcgatcaagcc | 53-tacatgtgaattcgtttcagcatc | 5′ of smu.299 for mutagenesis |
| smu.299 3F | 35-tagtaaatagctggaattcaggtgc | 33-tgaaaccagatcagctgttgac | 3′ of smu.299 for mutagenesis |
|
| |||
| Primers for qPCR | Forward (5′–3′) | Reverse (5′–3′) | Application |
|
| |||
| 16S-1 | rRNA cacaccgcccgtcacacc | cagccgcaccttccgatacg | 16S rRNA fragment, 160 bp |
| smu.1128 | aagggtgggttcggacttgg | gcaggcgagcttcaaacattac | ciaH fragment, 115 bp |
| smu.1528 | cggatgcgtgttgctcttactg | ggctgataaccaacggctgatg | atpD fragment, 164 bp |
| smu.1877 | ggtccttcacttggcata | ctgccattggtaagttcatccc | mannose-PST, EIIDMan, 79 bp |
| smu.1961 | ggaagccctttgacaacagc | gcattcatcaatctggttctat | levD fragment, 94 bp |
| smu.299 | acgatggagctaatggctatgc | agcgtaagcggcaaaacttg | smu.299 fragment, 140 bp |
| smu.423 | ggtggtggtatgattagatgtgc | ccagaccagcctcctaaagc | smu.423 fragment, 137 bp |
| smu.43 | agcaaccagttatcttagg | gtataatatagaatcccgaatagg | smu.43 fragment, 200 bp |
| smu.44 | tggcagactgggaaatataagc | ggcaaactcactcattgacaac | smu.44 fragment, 148 bp |
| smu.45 | gttacaggataccacggctgaag | accttgagttgccatagttcgtag | Smu.45 fragment, 96 bp |
| smu.46 | cacatagtgatgatgtccaaattg | gcatcagacttctttaaacttg | smu.46 fragment, 199 bp |
| smu.47 | ctgccacatagacgagaa | gtttacaattccacccacaa | smu.47 fragment, 124 bp |
| smu.610 | ttgccgatgaaacgaccactac | tcagcttccttactcgcactcc | spaP fragment, 115 bp |
| smu.78 | gggacttgggaagtacgagaag | aaacaagagctgctgcaccg | fruA fragment, 148 bp |
| smu.872 | gcggcttatgttacgggtacg | aaagcagtggtcgcaacaaaag | fruI fragment, 116 bp |
| smu.985 | cgccgtttatgtcaggaggtatc | ggcattggataagcaggcatagc | blgA fragment, 178 bp |
| smu.987 | acgactgcttctcaaacgaatg | ctgcttgttcacctgttgatgg | wapA fragment, 112 bp |
Kanr, Spr and Ermr, for kanamycin, spectinomycin and erythromycin resistance, respectively. Sequences underlined are restriction sites engineered for cloning.