TABLE 2.
PCR primers used in this study
| Name | Size (nt) | Sequence (5′→3′)a | Function of PCR product |
|---|---|---|---|
| 5DifCAT | 81 | CCTTAGGATGCATGGTGCGCATAATGTATATTATGTTAAATCCCTTAT GCGACTCCTGCATCCCTTTCGTCTTCGAATAAA | difE. coli-cat-difE. coli for pTOPO-DifCAT |
| 3DifCAT | 81 | CCTTAGGATGCATATTTAACATAATATACATTATGCGCACCATCCGCTT ATTATCACTTATTCAGGCGTAGCACCAGGCGT | |
| 5bac.DifCAT | 84 | AATACCGGTGGTGACCACTTCCTAGAATATATATTATGTAAACTTATT CTTCAACTAAAGCACCCATTAGTTCAACAAACGAAA | difB. subtilis-cat-difB. subtilis for pTOPO.bac-DifCAT |
| 3bac.DifCAT | 77 | TTGAATATTAGTTTACATAATATATATTCTAGGAAGTGATATCTTCAAC TAACGGGGCAGGTTAGTGACATTAGAAA | |
| msb.int F | 70 | TGCGGCGAAAACGCCACATCCGGCCTACAGTTCAATGATAGTTCA ACAGAAGTGTGCTGGAATTCGCCCT | msbB deletion |
| msb.int R | 70 | TTGGTGCGGGGCAAGTTGCGCCGCTACACTATCACCAGATTGATT TTTGCATCTGCAGAATTCGCCCTTA | |
| Int F | 70 | AAACCCGCCCCTGACAGGCGGGAAGAACGGCAACTAAACTGTTAT TCAGTTTGCGCCGACATCATAACGG | rbpA insertion |
| Int R | 70 | GCCGGATGCGGCGTGAACGCCTTATCCGGTCTACCGATCCGGCAC CAATGGCTACGGTTTGATTAGGGAA | |
| SEQ5MPR | 20 | TGTTGAAGGATTGGAAAACG | mpr cloning locus |
| SEQ3MPR | 20 | ACTAATGGAATGGCATGATC | |
| YKTD | 21 | AATTAGAGACGTTAAGCTGGA | nprE cloning locus |
| SEQ3NPRE | 20 | ATACATAATGACTGAATAAC | |
| SML | 20 | TGACCTGGTGATTGTCACCC | msbB screening locus |
| SMR | 20 | TAAACCAGCAGGCCGTAAAC | |
| UbiB F | 20 | GATCGCCTGTTTGGCGATGC | ubiB-FadR screening locus |
| UbiB R | 20 | GAATCTGATGGAACGCAAAG | |
| C5MPR | 20 | TTTCGAATCAGAAATCACAC | mpr screening locus |
| C3MPR | 20 | CTACTCTTTCAGGCGCGCGG | |
| C5NPRE | 20 | TGATCAACCTCGAAAACCTG | nprE screening locus |
| C3NPRE | 20 | GTATATGGCATTACTGCACC |
a
For primers 5DifCAT and 3DifCAT, the difE. coli sites are double underlined and the cat homology regions are boldfaced. For primers 5bac.DifCAT and 3bac.DifCAT, the difB. subtilis sites are double underlined and the cat homology regions are boldfaced. For primers msb.int F and msb.int R, the homology regions flanking msbB are underlined and the pTOPO-DifCAT homology regions are boldfaced. For primers Int F and Int R, the homology regions flanking ubiB/fadR are underlined and the prbpA-DifCAT homology regions are boldfaced.