TABLE 4.
Primers used in this study
| Primer | Sequence (5′ to 3′)a | Purpose |
|---|---|---|
| 1004uF | CTATGACATGATTACGAATTCACGATTTGCCTTATTACCACC (EcoRI) | Amplification of upstream flanking region of bmoA for construction of pK18mobsacB::ΔbmoA |
| 1004uR | GGATCCCCTTCATTGCACATCGTTCA (BamHI) | |
| 1004dF | GGATCCGAGCAGATCCATGAAGGCAT (BamHI) | Amplification of downstream flanking region of bmoA for construction of pK18mobsacB::ΔbmoA |
| 1004dR | ACGACGGCCAGTGCCAAGCTTCCTCGTGATAGTGGCTCATG (HindIII) | |
| 1005uF | CTATGACATGATTACGAATTCCCACCTTGACGTTATCCATC (EcoRI) | Amplification of upstream flanking region of bmoB for construction of pK18mobsacB::ΔbmoB |
| 1005uR | CATGACTCTTGAAGAAATTCTGGGTCATGGA | |
| 1005dF | GAATTTCTTCAAGAGTCATGTCGCCATCG | Amplification of downstream flanking region of bmoB for construction of pK18mobsacB::ΔbmoB |
| 1005dR | ACGACGGCCAGTGCCAAGCTTTGCCATCGTGTATGTGTCA (HindIII) | |
| sacB-F | CGACAACCATACGCTGAGAG | Amplification of partial sacB gene fragment |
| sacB-R | CGAAGCCCAACCTTTCATA | |
| 1004-test-F | TGAACGATGTGCAATGAAGG | Primers for screening of bmoA double-crossover mutant strains |
| 1004-test-R | GCATACCCCATTCAATGTCA | |
| 1005-test-F | TTCGTAGGTCCTGGAGTAGG | Primers for screening of bmoB double-crossover mutant strains |
| 1005-test-R | CGATGGCGACATGACT | |
| 04HF | GTCGACGGTATCGATAGGTATCCTCGCTCGGGCATT | Amplification of bmoA for construction of pBBR1MCS-bmoA |
| 04HR | CGCTCTAGAACTAGTGGAAAGTCTACCTGGTCGC | |
| 05HF | GTCGACGGTATCGATTGAATGGGGTATGCGTGA | Amplification of bmoB for construction of pBBR1MCS-bmoB |
| 05HR | CGCTCTAGAACTAGTAGTCACGCTGAGCGCCC | |
| 04EF | CATGCCATGGATCTGCTCGCCACAT (NcoI) | Amplification of bmoA gene for construction of pET28a-bmoA |
| 04ER | CCCAAGCTTGCCCTGAACGATGTGCAA (HindIII) | |
| 05EF | CGCCATATGACCCAGAATTTCTTCAATC (NdeI) | Amplification of bmoB gene for construction of pET28a-bmoB |
| 05ER | CGCGGATCCTCAGAACTCGATGGCGAC (BamHI) | |
| 04EF2 | CATGCCATGGATCTGCTCGCCACAT (NcoI) | Amplification of bmoA gene for construction of pACYC-bmoA |
| 04ER2 | CCCAAGCTTTCAGCCCTGAACGATGTG (HindIII) | |
| 05EF2 | CGCCATATGACCCAGAATTTCTTCAATC (NdeI) | Amplification of bmoB gene for construction of pACYC-bmoB |
| 05ER2 | TCGCGATCGTCAGAACTCGATGGCGAC (PvuI) |
a
Restriction sites are underlined and the restriction enzymes are in parentheses next to the sequence.