From: GTP-binding protein Era: a novel gene target for biofuel production
 | Description | Reference |
---|---|---|
Strain | Â | Â |
E. coli DH5α | Used for molecular cloning | [25] |
S. elongatus PCC 7942 | Wild-type freshwater cyanobacterium | gifted by Susan S. Golden (UCSD, California) |
2A01 | S. elongatus PCC 7942 containing transposon pRL1063a | This study |
PCC 7942 | S. elongatus PCC 7942 containing pNS3 | This study |
Se:era | S. elongatus PCC 7942 expressing the era gene from neutral site 3 | This study |
Se:Δera | S. elongatus PCC 7942 with a insertion of the inactivate transposon (pSE3) and expressing the era gene from neutral site 3 | This study |
Se:Δera + era | S. elongatus PCC 7942 expressing a truncated thioesterase from E. coli DH5α ('tesA) and containing a disrupted acyl-ACP synthetase (SynPCC 7942_0918) | This study |
Plasmid | Â | Â |
pJet1.2 | Used for the cloning of blunt PCR products | Thermo-Fisher Scientific (Waltham, MA, USA) |
pRL1063a | Contains a transposon based on Tn5, which bears several antibiotic resistance genes (kanamycin (Km), bleomycin (Ble) and streptomycin (Sm)), promoterless luciferase (luxAB) reporter genes, an oriV not recognized by PCC 7942 and a transposase gene. The oriT for conjugative transfer is present on the plasmid but is not part of the transposon. | [11] |
pHN1-LacUV5 | Targets Neutral site 3, confers resistance to chloramphenicol antibiotic (Cm) and contains a strong isopropyl-β-D-thiogalactopyranoside (IPTG)-regulated lacUV5 promoter followed by a ribosome binding site (RBS) and an unique HindIII restriction site | [15] |
pNS3 | Derived from pHN1-LacUV5 with the original RBS removed and lacking the ATG start codon near the multiple cloning site | This study |
pNS3:ERA | Derived from pNS3 with era gene (Synpcc7942_0160) cloned into the unique HindIII restriction site behind the inducible lacUV5 promoter | This study |
pSe:Δera | Inactivate transposon TN5-1063a originally isolated from strain 2A01, with the transposase mutate by digestion with NotI and blunting using T4 DNA polymerase | This study |