TABLE 2.
Primers used in this study
| Primer | Sequencea | Characteristic/function |
|---|---|---|
| P-for-nisA-T | GGAAGATCTAGTCTTATAACTATAC | BglII cleavage site |
| P-rev-nisA-T-2nd | CGGGGTACCTGTTTTTTCCTCTC | KpnI cleavage site |
| P-for-KpnI-ltnJ | CGGGGTACCCTGTAAGGAGAAAAATTATG | KpnI cleavage site |
| P-rev-BglII-Cmr | GGAAGATCTTGGAGCTGTAATATAAAAAC | BglII cleavage site |
| P-for-HindIII-T-ltnJ | CCCAAGCTTGTAAGCAAATAACCAAATC | HindIII cleavage site |
| P-ltnJ-rev-XhoI | CCGCTCGAGTTAGTGGTGGTGGTGGTGGTGTGTATCATAAGAAGTATCATATCTC | XhoI cleavage site |
| P-for-ALTIK | GCGTTAACAATTAAATAAGCTTTCTTTGAACC | General forward primer; 5′ phosphorylation |
| P-rev01 | AAGAGATATAGCTGCTTTCATGTTACAACCCATC | Construction of NisAtail |
| P-rev02 | AAGAGAAGATATAGCTGCTTTCATGTTACAACCCATC | Construction of S26insS |
| P-rev03 | AAGAGAAGAAGATATAGCTGCTTTCATGTTACAACCCATC | Construction of S26insSS |
| P-rev04 | AAGTGTAGATATAGCTGCTTTCATGTTACAACCCATC | Construction of S26insT |
| P-rev05 | TTTAGATATAGCTGCTTTCATGTTACAACCCATC | Construction of L27K |
| P-rev06 | AAGAGATTTAGCTGCTTTCATGTTACAACCCATC | Construction of I25K |
| P-rev07 | TTCAGATATAGCTGCTTTCATGTTACAACCCATC | Construction of L27E |
| P-rev08 | AAGAGATTCAGCTGCTTTCATGTTACAACCCATC | Construction of I25E |
| P-rev09 | AAAAGATATAGCTGCTTTCATGTTACAACCCATC | Construction of L27F |
| P-rev10 | AAGAGAAAAAGCTGCTTTCATGTTACAACCCATC | Construction of I25F |
| P-rev11 | ATTAGATATAGCTGCTTTCATGTTACAACCCATC | Construction of L27N |
| P-rev12 | AAGAGAATTAGCTGCTTTCATGTTACAACCCATC | Construction of I25N |
| P-rev13 | ACCAGATATAGCTGCTTTCATGTTACAACCCATC | Construction of L27G |
| P-rev14 | ACCAGAACCAGCTGCTTTCATGTTACAACCCATC | Construction of I25G-L27G |
| P-rev15 | AAGAGAACCAGCTGCTTTCATGTTACAACCCATC | Construction of I25G |
| P-rev16 | TGCAGATATAGCTGCTTTCATGTTACAACCCATC | Construction of L27A |
| P-rev17 | TGCAGATGCAGCTGCTTTCATGTTACAACCCATC | Construction of I25A-L27A |
| P-rev18 | AAGAGATGCAGCTGCTTTCATGTTACAACCCATC | Construction of I25A |
| P-rev19 | AAGTGTTATAGCTGCTTTCATGTTACAACCCATC | Construction of S26T |
| P-for-K34L | TTTTAAGCTTTCTTTGAACC | Construction of nisin(Δ30–34)-KIHIHVSL |
| P-rev-K34L | GCTTACGTGAATACTACAATG | Construction of nisin(Δ30–34)-KIHIHVSL; 5′ phosphorylation |
| P-for-S29insKIH | AAAATTCACATTCACGTAAGCTTTTAAGC | Construction of nisin(Δ30–34)-KIHIHVSL |
| P-rev-S29insKIH | ACTACAATGACAAGTTGCTG | Construction of nisin(Δ30–34)-KIHIHVSL; 5′ phosphorylation |
a
Restriction sites engineered in the primers are underlined.