TABLE 1.
Strains and plasmids
| Strain or plasmid | Characteristics or genotype |
|---|---|
| E. coli strains | |
| AM1452 | HB101 containing helper plasmid pRL528a and transposon plasmid pAM1037 |
| AM1460 | HB101 containing plasmid pRK2013b to provide conjugal transfer functions |
| DH10B | Host for plasmids |
| Synechococcus strainsc | |
| PCC 7942 | Wild-type Synechococcus |
| AMC149 | psbAI::luxAB reporter gene fusion inserted into NSId |
| AMC393 | psbAI::luxAB reporter gene fusion inserted into NSIIe (Cmr) psbAI::luxCDE fusion inserted into NSI (Spr) |
| AMC408 | purF::luxAB reporter gene fusion inserted into NSIIe (Cmr) psbAI::luxCDE fusion inserted into NSI (Spr) |
| AMC412 | Derivative of AMC149 with psbAI::luxCDE fusion inserted into NSII (from pAM1706) |
| AMC412(cpmA::Km) | Derivative of AMC412 in which cpmA has been inactivated by recombination with pAM2087 (Kmr) |
| AMC462 | kaiBC::luxAB reporter gene fusion inserted into NSI |
| AMC509 | kaiA::luxAB reporter gene fusion (translational) inserted into NSI |
| AMC520 | psbAII::luxAB reporter gene fusion (translational) inserted into NSI |
| AMC537 | psbAIII::luxAB reporter gene fusion inserted into NSI |
| AMC538 | rpoD2::luxAB reporter gene fusion inserted into NSI |
| AMC539 | conII::luxAB reporter gene fusion inserted into NSI |
| AMC540 | Derivative of AMC149 with psbAI::luxCDE fusion and an ectopic copy of cpmA inserted into NSII (from pAM2089) |
| AMC540(cpmA::Km) | Derivative of AMC540 in which cpmA has been inactivated by recombination with pAM2087 (Kmr) |
| AMC543 | psbAI::luxAB reporter gene fusion inserted at BstEII site (Kmr selection) and psbAI::luxCDE inserted at BglII site (Cmr selection) of NSII |
| AMC550 | Derivative of AMC543 in which an IPTG-inducible ectopic copy of cpmA is inserted at NSI (from pAM1850) |
| LUX6 | ndhD::luxAB reporter gene fusion; a 3.0-kb genomic DNA fragment including ndhD is integrated into the ndhD locus by single recombination |
| Plasmids | |
| pAM1037e | Tn5 plasmid derivative pRL1058, further modified by insertion of a 0.7-kb fragment in the XbaI site to add outward-reading promoters from the Anabaena sp. strain PCC 7120 glnA and rbcL genes |
| pAM1706 | Vector that transfers psbAI::luxCDE fusion to NSII (subsite NS2.1, BstEII) of the Synechococcus chromosome |
| pAM1825 | NSI vector that carries lacIq, IPTG-inducible trc promoter, and ribosome-binding site upstream of unique NcoI and SmaI cloning sites |
| pAM1850 | Vector that transfers psbAI::luxCDE fusion to NSII (subsite NS2.2, BglII) of the Synechococcus chromosome |
| pAM2086 | pUC18 containing 0.9-kb EcoRI fragment including cpmA ORF in the same orientation as the lacZ promoter |
| pAM2087 | 46-bp NruI fragment of pAM2086 was replaced by a Kmr cassette |
| pAM2088 | 46-bp NruI fragment of pAM2086 was replaced by an Spr-Smr cassette |
| pAM2089 | Derivative of pAM1706 that carries a PlacZ::cpmA fusion downstream of psbAI::luxCDE |
| pAM2107 | Entire cpmA ORF inserted into pBR322 at the EcoRV site |
| pAM2108 | 0.9-kb EcoRI fragment from pAM2107 (with ends modified as described in Materials and Methods) inserted into pAM1825 for IPTG-inducible expression of cpmA |
a
Reference 8.
b
Reference 7.
c
All Synechococcus strains carry a psbAI::luxCDE fusion in NSII except PCC 7942, AMC149, AMC393, and AMC408; all except PCC 7942 and AMC149 are bioluminescent without exogenously added aldehyde substrate. All insertions into NSI were selected by Spr; all insertions into NSII were selected by Cmr, except as otherwise noted for AMC543 and ACM550.
d
Reference 19.
e
Reference 1.