TABLE 2.
Synthetic oligonucleotides used for targeted deletion or disruption and confirmation of insertion
| Target | Gene IDa | Type of inactivation | Oligonucleotide and sequence (5′ to 3′)b |
|---|---|---|---|
| comGB | 1029176 | Disruption | FP042, AACAGCTTATTATGCGCGTGAATG |
| FP043, AAAGTGTTCCCATGTTTCCTCAGC | |||
| comX | 1029188 | Disruption | FP044, GTGAAGATTGGCAACAAGAGG |
| FP045, TCACATGTTCCATTCTTTCCC | |||
| comE | 1029114 | Disruption | FP053, GACGTCTTGAAACCACCATTGCA |
| FP054, TGCCGTAGAATTCAATCCGTTCA | |||
| comC | 1029111 | Deletion | FP009, TATGGACCAAGAAATGCTGT |
| FP010, AGG/CGCGCCGATAATCTCTAATTCATCAGTCTT | |||
| FP011, AGGCCGG/CCAGAAGTTTTACACAAGCTTTG | |||
| FP012, TGGAGTTGCTTGATTTCATT | |||
| comD | 1029115 | Deletion | FP053, GACGTCTTGAAACCACCATTGCA |
| FP013, AGG/CGCGCCCGTAACAGCAATCATTATCAGAAA | |||
| FP014, AGGCCGG/CCAATCGCGGTATTGGTTTAGCA | |||
| FP012, TGGAGTTGCTTGATTTCATT | |||
| Kan cassette | FP001, AGG/CGCGCCGTTTGATTTTTAATG | ||
| FP002, AGGCCGG/CCATCGATACAAATTCCTC | |||
| FP037, TCATTTTCTCCCACCAGCTT; confirm insertion | |||
| FP038, GCGCCTACGAGGAATTTGTA; confirm insertion | |||
| pSF151 | FP039, AGCGGATAACAATTTCACACAGGA; confirm insertion |
a
S. mutans UA159 gene identification number (ID).
b
Restriction sites are in boldface: AscI, GG/CGCGCC; and FseI, GGCCGG/CC.