TABLE 2.
Oligonucleotides used in this study
| Use and oligonucleotide | Sequence | Descriptiona |
|---|---|---|
| Amplification | ||
| TSp26 | CATCGAACTTTGACATATTG | |
| TSp27 | GGTATCGATAAGCTTGATATCGAATTCCTGCAGCCCGGGGGAAGACATTACCGTAGATGG | |
| TSp28 | GTCTATTGCAAACTTGCTTC | |
| TSp29 | AGCTCCACCGCGGTGGCGGCCGCTCTAGAACTAGTGGATCCCATTGTTTCAGATGTATCG | |
| TSp30 | AGGTATTGAAAGGAGGTGACCAATATGTCAAAGTTCGATGGGATCGATCTGTATAATAAAG | |
| TSp31 | TTACAAGTGATTGTTTGAAGGAAGCAAGTTTGCAATAGACGTTTATAAGTGGGTAAACCGTG | |
| BS53 | GGCCAGTGAGCGCGCGTAATACGACTCACTATAGGGCGAACCATCATTGATGGTTTCTTTC | |
| BS54 | GATTACGCCAAGCGCGCAATTAACCCTCACTAAAGGGAACAAGCGGAAGAATGAAGTAAG | |
| TSp19 | TTCAGGTGCTTTTTTTATTTTATAAACTCATTCCCTGATCGCCTAAAATGTTAACACTTC | |
| TSp45 | TCATCATCGCTCATCCATGTCGACGGTATCGATAAGCTTG CATGGTTACAGCGGTATCGGTC | |
| CG1 | ATTGGTCACCTCCTTTCAATAC | |
| CG2 | GATATTTGTCTGTTACTATTTAGGTATTGAAAGGAGGTGACCAATGGATCCGTAAAACCATTAGCATCACCTTGC | |
| NN16 | TCAGCCATGGGATCCCCAGCTTGTTGATACACTA | |
| NN17 | TCAGAAGCTTGGCAGACATGGCCTGCCCG | |
| In vitro mutagenesis | ||
| TSp48 | CTCCGCAAATTAAAAGTGAATCAC | fw W1I |
| TSp49 | GTGATTCACTTTTAATTTGCGGAG | rv W1I |
| TSp73 | CGCAAATTACAAGTGAATCAC | fw W1I/K2T |
| TSp74 | GTGATTCACTTGTAATTTGCG | rv W1I/K2T |
| TSp75 | GCAAATTACAAGTATTTCACTTTGTACACC | fw W1I/K2T/E4I |
| TSp76 | GGTGTACAAAGTGAAATACTTGTAATTTGC | rv W1I/K2T/E4I |
| TSp76_1 | GGATGTAAAACTGGTGCATTGC | fw W1I/K2T/E4I/V12K |
| TSp76_i | GCAATGCACCAGTTTTACATCC | rv W1I/K2T/E4I/V12K |
| TSp69 | GGTGCATTGATGACTTGCTTCCTTC | fw Q17 M |
| TSp70 | GAAGGAAGCAAGTCATCAATGCACC | rv Q17 M |
| Gap repair | ||
| CG11 | ACTTGCTTCCTTCAAACACTAACTTGTAACTGCAAAATCTCTAAATAAGTAAAACCATTAGCATCACCTTGCTCTGACTCCTTGCACT | Universal oligonucleotide |
| CG12 | TTATTTAGAGATTTTGCAGTTACAAGTTAGTGTTTGAAGGAAGCAAGTTTGCAATGCACCAGTTACACATCCTGGTGTACAAAGTGATTCACTTTT | Universal oligonucleotide |
| TSp191 | TGTGAAAGTCTCTAAACAAGACTCAAAAATCACTCCGCAAGCAAAAAGTGAATCACTTTGTACACCAGGATGTGTAACTGGTGCATTGCAA | W1A |
| TSp192 | TGTGAAAGTCTCTAAACAAGACTCAAAAATCACTCCGCAATTCAAAAGTGAATCACTTTGTACACCAGGATGTGTAACTGGTGCATTGCAA | W1F |
| TSp193 | TGTGAAAGTCTCTAAACAAGACTCAAAAATCACTCCGCAATATAAAAGTGAATCACTTTGTACACCAGGATGTGTAACTGGTGCATTGCAA | W1Y |
| TSp194 | TGTGAAAGTCTCTAAACAAGACTCAAAAATCACTCCGCAAGATAAAAGTGAATCACTTTGTACACCAGGATGTGTAACTGGTGCATTGCAA | W1D |
| TSp195 | TGTGAAAGTCTCTAAACAAGACTCAAAAATCACTCCGCAAGAGAAAAGTGAATCACTTTGTACACCAGGATGTGTAACTGGTGCATTGCAA | W1E |
| TSp196 | TGTGAAAGTCTCTAAACAAGACTCAAAAATCACTCCGCAACATAAAAGTGAATCACTTTGTACACCAGGATGTGTAACTGGTGCATTGCAA | W1H |
| TSp197 | TGTGAAAGTCTCTAAACAAGACTCAAAAATCACTCCGCAAAAAAAAAGTGAATCACTTTGTACACCAGGATGTGTAACTGGTGCATTGCAA | W1K |
| TSp198 | TGTGAAAGTCTCTAAACAAGACTCAAAAATCACTCCGCAAAGAAAAAGTGAATCACTTTGTACACCAGGATGTGTAACTGGTGCATTGCAA | W1R |
| TSp208 | TGTGAAAGTCTCTAAACAAGACTCAAAAATCACTCCGCAATGGACAAGTATTTCACTTTGTACACCAGGATGTAAAACTGGTGCATTGATG | K2T/E4I/V12K/Q17 M |
| TSp209 | TTATTTAGAGATTTTGCAGTTACAAGTTAGTGTTTGAAGGAAGCAAGTCATCAATGCACCAGTTTTACATCCTGGTGTACAAAGTGAAATACTTGT | K2T/E4I/V12K/Q17 M |
| TSp225 | TGTGAAAGTCTCTAAACAAGACTCAAAAATCACTCCGCAACTTAAAAGTGAATCACTTTGTACACCAGGATGTGTAACTGGTGCATTGCAA | W1L |
| TSp226 | TGTGAAAGTCTCTAAACAAGACTCAAAAATCACTCCGCAAGTAAAAAGTGAATCACTTTGTACACCAGGATGTGTAACTGGTGCATTGCAA | W1V |
a
Letters indicate the amino acid residues exchanged (corresponding to the IUPAC abbreviation), and numbers indicate the positions of the exchanged amino acids in the respective subtilin variants. fw, forward; rv, reverse.