TABLE 3.
List of primers used in this study
| Primer name by application | Sequence (5′-3′)a | Restriction site |
|---|---|---|
| For gene knock out | ||
| 400upstream-F | ACACAGAGCTCAGCTGGTGAAATGGATGGC | SacI |
| 400upstream-R | TTAACTTTTCTCCTTAAAATAACCAAAATCTTTTCTT | |
| 400downstream-F | TTGGTTATTTTAAGGAGAAAAGTTAAAAGTATG | |
| 400downstream-R | ACCAAGCTAGCTGTATCACTTTTGGTATTGA | NheI |
| Seq1-F | ATTGGGTTCTAACAGAATGCGT | |
| INTZ-R | TTCTCCCCCTAATAATTCTGCAG | |
| INTZ-F | GGTTTTTATATTACAGCTCCAAG | |
| Seq1-R | AATCCTGAATCACTTTCGGTTT | |
| 1778upstream-F | CAACGGAGCTCCGAAACTAATGGCATCAAT | SacI |
| 1778upstream-R | GTTTTTGGTGCAGAGAAATTTATATATAAAATAACGATTTTGT | |
| 1778downstream-F | TTGTATAGGAGAAACAAGGTTTCTTGATAATAAATAACATTAATAGC | |
| 1778downstream-R | CTTTAAGGATCCAAAAGAAGAAGCTAGAAAGGCTT | BamHI |
| Seq2-F | CACATAATACCAGCAGTCAACGAA | |
| Seq2-R | GTGCTCCAAGTAGTATCATAGCGAT | |
| 1709upstream-F | TTCCTGGAGCTCAAACTTTATTTTGTTCTGCCAA | SacI |
| 1709upstream-R | ATGTTATTTATTATCAAGAAACCTTGTTTCTCCTATACAATGT | |
| 1709downstream-F | TTGTATAGGAGAAACAAGGTTTCTTGATAATAAATAACATTAATAGC | |
| 1709downstream-R | CTTTAAGGATCCAAAAGAAGAAGCTAGAAAGGCTT | BamHI |
| Seq3-F | GGCTAAAAACAGACTCCATCAATC | |
| Seq3-R | AGCCCGTTCTTTCTTATAACTTTAA | |
| For complementation | ||
| EPS-Promoter-F | AATTAGGTACCACACTGTAAAAATAAATAAGATCCT | KpnI |
| EPS-Promoter-R | TTAACCTCTTGTGCCATCATTTTATTCCTCTTTTATTTTT | |
| N506_0400-F | AAAAATAAAAGAGGAATAAAATGATGGCACAAGAGGTTAA | |
| N506_0400-R | CTTTTAAGCTTAATACGCACTATTTGGATGAAT | HindIII |
| N506_1778-F | TACATCGGTACCAAAATATGCGGTATGTATTTATCG | KpnI |
| N506_1778-R | GGTGCAGGATCCTTAATTCTTTTTCTTTCGTTTAAGTT | BamHI |
| N506_1709-F1 | AAGACAGATCTCGTAATTAAATTGATCAAGTACATTAT | BglII |
| N506_1709-R1 | ACAAAGGTACCACCAGATTCCAT | KpnI |
| N506_1709-F2 | CTGGTGGTACCTTTGTTTCAAAAGT | KpnI |
| N506_1709-R2 | ATTATGAGCTCTTATCTAATTCGGTGTTTTCTTCTACTT | SacI |
| For overexpression of N506_1709 in L. lactis | ||
| N506_1709Re-F | GAGAAACCATGGATGCTATCTAAAAATAATTTTCATG | NcoI |
| N506_1709 Re-R | TTAATGAGCTCTTACGCTTCCGGTTCTCTAATTCGGTGTTTTCTTCTACTT | SacI |
| For qRT-PCR | ||
| L. gasseri 16S-F | ACCCTTGTCATTAGTTGCCATCA | |
| L. gasseri 16S-R | GCTTCTCGTTGTACCGTCCATT | |
| 1709-F | ATTGGAACGATTTGAAGAGCG | |
| 1709-R | GAATCAGTAGTGTGGGAACCGAC | |
| L. lactis 16S-F | TCGTGTCGTGAGATGTTGGGT | |
| L. lactis 16S-R | GTCATAAGGGGCATGATGATTTG |
a
The restriction site is underlined in the primer sequence.