TABLE 1.
PCR primers used
| Role | Primer no. | Name | Sequence (5′-3′) |
|---|---|---|---|
| Recombinant sigma factor expression | 1 | SigA-GW-F | CTGGTTCCGCGTGGATCCAAAGCAAGCACGGCGAC |
| 2 | SigA-GW-R | CAAGAAAGCTGGGTCTCAGTCCAGGTAGTCGCGCAGG | |
| 3 | SigF-GW-F | CTGGTTCCGCGTGGATCCGGCGGTTCTGCATCGC | |
| 4 | SigF-GW-R | CAAGAAAGCTGGGTCCTACTCCAACTGATCCCGTAGCCGTG | |
| 5 | attB1-thrombin | GGGGACAAGTTTGTACAAAAAAGCAGGCTTCCTGGTTCCGCGTGGATC | |
| 6 | attB2 | GGGGACCACTTTGTACAAGAAAGCTGGGTC | |
| Real-time in vitro transcription template | 7 | F-rrnABt1(PlacZ) | TAGACTGAGCTCTCGAAGCTTATCGATGTCGACGTAG |
| 8 | R-rrnABt1 | AGTCAGTCGAGCTCAGAGATTTTGAGACACAACGTCGC | |
| 9 | F-PrsbW | CGGCGTCGGAGCTCTTGC | |
| 10 | R-PrsbW | CTAGTTAACTACGTCGACATCGATAAGCTTCGATCGACAGTACTCCCG | |
| 11 | F-rrnABt1(PrsbW) | CGGGAGTACTGTCGATCGAAGCTTATCGATGTCGACGTAGTTAACTAG | |
| 12 | BsiWI-polyA | GTACAAAAAAAAAAAAAAAAAAAAAAAAA | |
| 13 | BsiWI-polyT | GTACTTTTTTTTTTTTTTTTTTTTTTTTT | |
| Radiometric in vitro transcription template | 14 | IVT PCR F-Hsp60 | AATACCAGCCAGACGAGACGG |
| 15 | IVT PCR R-Hsp60 | AACCCCGCTACTTACGCTGG | |
| 16 | PrsbW IVT temp F | ATTGGCCTGGGTATCTGTTG | |
| 17 | PrsbW IVT temp R | TTGGTGGGTAAATCCGAGTC | |
| Pristinamycin-inducible SigF plasmid | 18 | F pri HindIII/Pptr | AGTCACAAGCTTACGAGTTCCGAGATCACCCCG |
| 19 | R pri Pptr/sigF | GGCAGCGCGCGCCGTCACTCAGGCTCCTTGTACGGTGTACGGG | |
| 20 | F Pri Pptr/sigF | CCCGTACACCGTACAAGGAGCCTGAGTGACGGCGCGCGCTG | |
| 21 | R Pri SigF/HindIII | AGTCACAAGCTTCTACTCCAACTGATCCCGTAGCCGT | |
| SigF knockout cassette | 22 | rsbwFKO(1)-F | CCCAGCGGATCCTGCTGG |
| 23 | rsbwKO(1)-R | CTGTGAGATACACTAGTGCGCAGCAGCGCC | |
| 24 | sigFKO(2)-F | GCTGCGCACTAGTGTATCTCACAGATGCACGTGTCG | |
| 25 | sigFKO(2)-R | CACCGAGGATCCCACCCC | |
| 26 | speI-HYG-F | GTACTAACTAGTCTGAGCTTGCATGCCTGC | |
| 27 | HYG-SpeI-R2 | TAACTAGTGACCACCCTGGAGGAGATGAT | |
| 28 | KO check F | GACCAGCTCGAAGACCAGAC | |
| 29 | KO check R | GCTCGCCGAGATCAAGTAAG |