TABLE 3.
Sequences of primers used in this study
| Primer | Sequence (5′ to 3′)a | Purpose |
|---|---|---|
| accBCPgapAFusF | CCTACAATCTTTAGAGGAGACACAACGTGTCAGTCGAGACTAGGAAGATCACCAAG | Expression of accBC |
| accBCdown100RKpnI | CTTGGTACCGAAATCTTGTTGTCGAATG | Expression of accBC |
| PgapAKpBgF | GCGGGTACCAGATCTGAAGATTCCTGATACAAATTCTGTTG | Expression of accBC and accD1 |
| accBCPgapAFusR | CTTGGTGATCTTCCTAGTCTCGACTGACACGTTGTGTCTCCTCTAAAGATTGTAGG | Expression of accBC |
| accD1PgapAFusF | CCTACAATCTTTAGAGGAGACACAACATGACCATTTCCTCACCTTTGATTGACGTC | Expression of accD1 |
| accD1down150RKpnI | TCGGGTACCGGTTATATTAGCCCAGCG | Expression of accD1 |
| accD1PgapAFusR | GACGTCAATCAAAGGTGAGGAAATGGTCATGTTGTGTCTCCTCTAAAGATTGTAGG | Expression of accD1 |
| InFu-accBCf | TGAGTCATCAATTTAAATCAGGAGTTATTAATGTCAGTCGAGACTAGGAAG | Expression of accBC, accD1, and accE |
| InFu-accBCr | TTACTTGATCTCGAGGAGAACAACGC | Expression of accBC, accD1, and accE |
| InFu-accEf | CTCGAGATCAAGTAAAAACTGTTTTTTAAAGGAGAACCATGTCTGAAG | Expression of accBC, accD1, and accE |
| InFu-accEr | TATGGATTCGCCGATCTAGAAGAAATTCACATTCTGAAACGCGC | Expression of accBC, accD1, and accE |
| InFu-accD1f | ATCGGCGAATCCATAAAGGTTCAAAAG | Expression of accBC, accD1, and accE |
| InFu-accD1r | TAAATTGATGACTCATTACAGTGGCATGTTGCCGTGCTTG | Expression of accBC, accD1, and accE |
| fasAexpFupEcoRV | TGGGATATCCTGTGGTGGCTTTTAAGAAG | Expression of fasA |
| fasAexpRdownNheI | TGCGCTAGCAAACTTGAGAAGTTTCATGAG | Expression of fasA |
| InVer-pCS299PfasAf | AAGTTTGCTAGCGCAGGCATGCAAGCTTGGCGTAATCATGG | Expression of fasA |
| InVer-pCS299PfasAr | CCACAGGATATCCCATGCAGGTCGACTCTAGAGGATCC | Expression of fasA |
| fasBexpFupBlnI | AGTCCTAGGCCGGGAGCTGTAGAAAATTGC | Expression of fasB |
| fasBexpRDownNheI | GTTGCTAGCACTAAGTTACCCTCGGTGTGAAG | Expression of fasB |
| InVer-pCS299PfasBf | CTTAGTGCTAGCAACGGCATGCAAGCTTGGCGTAATCATG | Expression of fasB |
| InVer-pCS299PfasBr | TCCCGGCCTAGGACTTGCAGGTCGACTCTAGAGGATCC | Expression of fasB |
| CglfasIA5′BglIIF | ACGAGATCTACGCATTCGTAAGTGG | Deletion of fasA |
| CglfasIAFusR | CAACGGATGCACGTGCCAGGAGGACGGTACCGGTTGCACGTGCCTTGGAAAC | Deletion of fasA |
| CglfasIAFusF | GTTTCCAAGGCACGTGCAACCGGTACCGTCCTCCTGGCACGTGCATCCGTTG | Deletion of fasA |
| CglfasIA3′BglIIR | CAGAGATCTTAGCTATCTAACGTTTAGC | Deletion of fasA |
| fasBupF | CAGTATTCCTGTGCATGTGAATACGC | Deletion of fasB |
| fasBFusR | AGGAGGACTGCAGCTTCAACTTCGTTCCTGCTCAATTCGGTCACGT | Deletion of fasB |
| fasBFusF | ACGTGACCGAATTGAGCAGGAACGAAGTTGAAGCTGCAGTCCTCCT | Deletion of fasB |
| fasBdnR | TCTTGATCAAGGTGCCGGTGGGAA | Deletion of fasB |
| ncrFBam2 | ACTGGATCCACACATAAGTGCTCT | Integration of bioI under gapA promoter into C. glutamicum genome |
| PgapAFusR | CTAAATTTCTTCCAACAAATCTTCCGTCTTGTTTCAGGCCACCACTTAGAAGGC | Integration of bioI under gapA promoter into C. glutamicum genome |
| PgapAFusF | GCCTTCTAAGTGGTGGCCTGAAACAAGACGGAAGATTTGTTGGAAGAAATTTAG | Integration of bioI under gapA promoter into C. glutamicum genome |
| bioIFusR | GCAGTTGACGATGCAATTGTCACGTTGTGTCTCCTCTAAAGATTGTAGG | Integration of bioI under gapA promoter into C. glutamicum genome |
| bioIFusF | CCTACAATCTTTAGAGGAGACACAACGTGACAATTGCATCGTCAACTGC | Integration of bioI under gapA promoter into C. glutamicum genome |
| bioIFusR2 | GACAATTGAATTACGCCCTAGTAGTAGATGTTCACTCCCCTTTTTTATAG | Integration of bioI under gapA promoter into C. glutamicum genome |
| ncrFusF | CTATAAAAAAGGGGAGTGAACATCTACTACTAGGGCGTAATTCAATTGTC | Integration of bioI under gapA promoter into C. glutamicum genome |
| ncrRBam2 | TACGGATCCCAGCATCATGCTTGT | Integration of bioI under gapA promoter into C. glutamicum genome |
a
KpnI sites are underlined, BglII sites are italicized, the BclI site is underlined and bold, and BamHI sites are in bold.