TABLE 1.
Primers used in this study
| Primer (relevant plasmid) | Sequencea |
|---|---|
| Amplification primers for gene inactivation fragments (pMUTIN2) | |
| mutyvqI+ | GACGCGGAATTCCGGCGGATTCTTATTAATTG |
| mutyvqI− | TTATACGGGATCCCGGAAGCCGGTTCAGGATC |
| mutyvqH+ | GACGCGGAATTCCCGATATGGAAAGCGACATCGC |
| mutyvqH− | TTATACGGGATCCCGCGCTTCCACATCGTCTGC |
| mutyvqG+ | GACGCGGAATTCCGAAGTGACATATCGGCAGGC |
| mutyvqG− | GACGCCGGGATCCCGGCTTACATTAGCATCACCGC |
| mutyvqF+ | GACGCGGAATTCCGCGTTTGCCGCATTTCTGATTTACGC |
| mutyvqF− | TTATACGGGATCCCGACACTTTTACCCGGCGC |
| mutyvqE+ | GACGCGGAATTCCAGCGTGACCGTCGGTTTCGC |
| mutyvqE− | GACGCCGGGATCCCGGTCCAAAAGCTCCGTAAGGC |
| mutyvcR+ | GACGCGGAATTCCGATTATGGGGCCGTCGGGAAGC |
| mutyvcR− | GACGCCGGGATCCCGGCGTAACCAACAAAATCGTCGC |
| mutyvcS+ | GACGCGGAATTCCCAGGCTGAAGGATCATGCACGC |
| mutyvcS− | GACGCCGGGATCCCGGTGACAGACTGTCAAAGC |
| mutyxaH+ | GACGCGGAATTCCGCCGCCGCAGACATACTG |
| mutyxaH− | TTATACGGGATCCCGGTGGATCCGCTGTTC |
| verifypMUTIN+ | ATAATTCTACACAGCCCAGTCCAGACTATTCGG |
| Amplification primers for gene overexpression (pHT315) | |
| pHTyvqIH+ | GACGCGGATCCAAAAGGAGAATGATAAAAATGAAAATAAACAAGAAAACAATAG |
| pHTyvqIH− | GACGCCCCGGGTTATTCATTTGCCGCTTTTGTCTGGTC |
| pHTyvqGFE+ | GACGCGGATCCAAAAGGAGAATGATAAAAATGGTCATTGAGTCGGATAGCAAGG |
| pHTyvqGFE− | GACGCCCCGGG TCAATCAATAATACTCGAATCACGTTC |
| pHTyvcRS+ | GACGCGGATCCAAAAGGAGAATGATAAAAATGAACGTGTTGCAAACAACGAACC |
| pHTyvcRS− | GACGCCCCGGGTTACATACGCTGAAGAACAGC |
| pHTyxaGH+ | GACGCCCCGGGAAAAGGAGAATGATAAAAATGAAAACATTATGTACACATTC |
| pHTyxaGH− | GACGCCCCGGGCTAGACTTTTGTTTTCTTTGCAAT |
| verifypHT+ | AGCGGATAACAATTTCACACAGGA |
| Amplification primers for nisin immunity genes and lacZ (pSac-Kan) | |
| nisFEG+ | TCGGAAGATCTTCTCCCCGCGGGGAGCCGGCGTTCGAAGGAACTACAAAATAAATTATAAGGAGGCACTCAAAATGCAGGTAAAAATTCAAAATCTTTCTAAAAC |
| nisFEG− | ATGCCCCTAGGTCTAATCTTTTTTTTAGATAATGCTACAAG |
| nisI+ | GACCTCTAGCTAGCTAGAGGAGGGAAGAGGAAAGTGGCCTTAATAGGGATAACAGGTTTATCAGG |
| nisI− | CGGGAAGATCTTCGACGTCGTTTCCTACCTTCGTTGCAAGCTTAAAATC |
| lacZ+ | ATGGCGGGGTACCCCTAACCTAACTAAAGGTGGTGAACTACTGTG |
| lacZ− | AGCTCCCTAGGGGTTATTATTATTTTTGACACC |
| Amplification primers for synthetic promoter in front of nisin immunity genes | |
| SP+ | ATGCGTCCCCGCGGGGATCCCCCGGGGGACAGTGATGGGTCCAGAAGGTGCGGCATCG |
| SP− | GCATGGCCGGCATTATANNNNNNNNNNNNNNNNNTGTCAANNNNNNNNNNNNNNNAAACGCAATATGATGCAGTCCCTGCCCTTTC |
a
Restriction sites are underlined.