TABLE 2.
Primers used in this study
| Function and primer name | Sequence (5′ to 3′)a | Restriction site |
|---|---|---|
| Deletion of divIB | ||
| divIB-F1 | GCTCTTGGCGATAGCAAAAT | |
| divIB-R2 | TCAAACAAATTTTCATCAAGCTTCTTTAGACTCTGGTTCTTCTTT | |
| divIB-F3 | AAAGAAGAACCAGAGTCTAAAGAAGCTTGATGAAAATTTGTTTGA | |
| divIB-R4 | GTCAATCGAATTAAATCGGATGTTCTAGAACTAGTGGATCCCCCGG | |
| divIB-F5 | CCGGGGGATCCACTAGTTCTAGAACATCCGATTTAATTCGATTGAC | |
| divIB-R6 | CCCCAAAAATTCACCATCTG | |
| Deletion of divIC | ||
| divIC-F1 | AGACCGTTTTGGAGAATACCC | |
| divIC-R2 | TCAAACAAATTTTCATCAAGCTTGTATTCATTTTGAATAAAAGAATT | |
| divIC-F3 | AATTCTTTTATTCAAAATGAATACAAGCTTGATGAAAATTTGTTTGA | |
| divIC-R4 | TTGATACTGAGTTTGCAAGTCTGTCTAGAACTAGTGGATCCCCCGG | |
| divIC-F5 | CCGGGGGATCCACTAGTTCTAGACAGACTTGCAAACTCAGTATCAA | |
| divIC-R6 | TCATTCGCATAAACAGGAACC | |
| Deletion of ftsL | ||
| ftsL-F1 | AAGTGAAGCCGATTGAGACAA | |
| ftsL-R2 | TCAAACAAATTTTCATCAAGCTTGTTTAAGTTGCATCTGTAGTAT | |
| ftsL-F3 | ATACTACAGATGCAACTTAAACAAGCTTGATGAAAATTTGTTTGA | |
| ftsL-R4 | CTTGCTTGGCATCGTCCAATTTCTAGAACTAGTGGATCCCCCGG | |
| ftsL-F5 | CCGGGGGATCCACTAGTTCTAGAAATTGGACGATGCCAAGCAAG | |
| ftsL-R6 | TTGTCCGTTTGGGTAACTACG | |
| Gene amplification for ligation into pCEP | ||
| divIB-D1 | CTGCCATGGCGTCAAAAGATAAGAAAAATGAGG | NcoI |
| divIB-R | CGCCCTCGAGGATCCACTAGTCTAGCGACGCGATGAACGC | BamHI |
| divIC-D1 | CTGCCATGGCGTCTAAAAATATTGTACAATTGAATAATTC | NcoI |
| divIC-R | CGCCCTCGAGGATCCACTAGTTCACCTTTGAAGCAAGTCAGG | BamHI |
| ftsL-D1 | CTGCCATGGCGGCAGAAAAAATGGAAAAAACAGG | NcoI |
| ftsL-R | CGCCCTCGAGGATCCACTAGTTTACTCCGCTATTCTAATATTTTCATTG | BamHI |
| Gene amplification for ligation into pET-30b(+) | ||
| divIB-F | GGTGCCATATGTCGACGTCAAAAGATAAGAAAAATGAGGACAAAGAAACC | NdeI |
| External primer | ||
| divIB-F0 | GAAAGAACTAACAGAGCGCTAC | |
| divIB-R7 | GATACCTGTTGCAAGTCCAGC | |
| amiF-F1 | CCGAATTAGCACGTTATCAAAAAGG | |
| treR-R | CGCCAAATCGCTGAGCAG | |
| Mutagenesis of divIB in pET30b-divIB | ||
| divIB E222A-F | GGTCAAGGCATATGATATTGTGGCC | |
| divIB E222A-R | GGCCACAATATCATATGCCTTGACC | |
| divIB Δγ-F | GGAAAAAGCCTAGCTAGCGGCCAAGG | NheI |
| divIB Δγ-R | CCTTGGCCGCTAGCTAGGCTTTTTCC | NheI |
| divIB S337*-F | GCCACAATTGTCAGAACCGTGAGTGGTCG | |
| divIB S337*-R | CGACCACTCACGGTTCTGACAATTGTGGC | |
| divIB Δβγ-F | CCAACTAAGTTCACTATCTAGATCAAGG | XbaI |
| divIB Δβγ-R | CCTTGATCTAGATAGTGAACTTAGTTGG | XbaI |
| divIB Δec-F | CAGTCCTTAATCGATCATGAAAGATATTCG | ClaI |
| divIB Δec-R | CGAATATCTTTCATGATCGATTAAGGACTG | ClaI |
a
Restriction sites are underlined.