Table 1.
Strains, plasmids, and primers used in this study. Restriction enzyme sites in the primers are underlined.
| Strain | Description | Reference |
|---|---|---|
| Escherichia coli DH5α | E. coli strain used for molecular cloning | New England Biolabs |
| Synechococcus elongatus PCC 7942 | Wild type; a freshwater cyanobacterium | ATCC |
| SE01 | S. elongatus 7942 with gene knockout of acyl-ACP synthetase (Synpcc7942_0918) | This study |
| SE02 | S. elongatus 7942 with gene knockout of acyl-ACP synthetase (Synpcc7942_0918) and expression of a truncated thioesterase from E. coli (‘tesA) | This study |
| Plasmids | Description | Reference |
| pAM2991 | Plasmid constructed for S. elongatus 7942 genome integration at NSI and gene expression using the trc promoter | (Mackey et al. 2008) |
| pSE15 | Plasmid derived from pAM2991 with NSI homologous regions replaced by upstream (961bp) and downstream (978bp) homologous regions of Synpcc7942_0918 for gene knockout of the acyl-ACP synthetase | This study |
| pSE16 | Plasmid derived from pSE15 with a truncated thioesterase from E. coli (‘tesA) expressed by the trc promoter | This study |
| Primers | Description |
|---|---|
| 5′-P-CTGGAGATCTGACGAGCAGGGACTCGAAGCT-3′ | Forward primer for removal of NSI 5′ region from pAM2991 |
| 5′-P-GTCACTCGAGCGGCTGCCGGATATCCTGCCT-3′ | Reverse primer for removal of NSI 5′ region from pAM2991 |
| 5′-P-CTAGCTTAAGACTCACCAGTCACAGAAAAGCATCT-3′ | Forward primer for removal of NSI 3′ region from pAM2991 |
| 5′-P-GTCCACTAGTATCTTCCTGCTCCAGAAGCTCGAAA-3′ | Reverse primer for removal of NSI 3′ region from pAM2991 |
| 5′-GTACTTCTCGAGGCAGCTCCGTTGTCGCAGTGTCAGA-3′ | Forward primer for cloning the region upstream of Synpcc7942_0918 |
| 5′-GAGTCGAGATCTGCCTGTGGTGCCCGAGGTATAGATC-3′ | Reverse primer for cloning the region upstream of Synpcc7942_0918 |
| 5′-GTCAGGACTAGTGAACCCCAGCCGATTGAAGATGCCT-3′ | Forward primer for cloning the region downstream of Synpcc7942_0918 |
| 5′-GAGTTGCTTAAGAGACATCACTCAAGTCATCAGTCACAG-3′ | Reverse primer for cloning the region downstream of Synpcc7942_0918 |
| 5′-GTGATGGAATTCGCAGCGGACACGTTATTGATTCTGG-3′ | Forward primer for cloning ‘tesA from E. coli DH5α |
| 5′-CGAGTCGGATCCTTATGAGTCATGATTTACTAAAGGCTGC-3′ | Reverse primer for cloning ‘tesA from E. coli DH5α |