Strains or Plasmids Strains Escherichia coli DH5a DH5alpir CC118lpir Ralstonia solanacearum AW1-AI8,395 Vibrio fischeri ES114 ANS3 ANS5 ANS6 ANS7 AMJ1 AMJ2 CL21 EVS102 JB33 NL11 VCW2G7 Plasmids pAJ4 pAJ7 pAKD601B pAKD702 pAS2 pAS3 pAS4 pAS6 pAS104 pBluescript pEVS104 pEVS122 pEVS148k pEVS149k pJLB169 pJLB171 pVSV102 pVSV208 Oligonucleotidesb ASind_arcAF ASind_arcAR ASLUX1 ASLUX2 F’/endA1 hsdR17 glnV44 thi-1 recA1 gyrA96 (NxR) relA1 D(lacIZYA-argF)U169deoR(f80dlacID(lacZ)M15) lpir derivative of DH5a D(ara-leu) araD Dlac74 galE galK phoA20 thi-1 rpsE rpsB argE(Am) recA lpir solI8::SP aidA395::Tn3HoHo1 HSL-deficient Lac+ NxR SpR CbR wild-type isolate from E. scolopes light organ ES114 DluxI; luxI 3OC6 synthase (VF_A0924) gene deletion ES114 DluxICDABEG ES114 DarcA::erm DluxICDABEG ES114 DarcA luxI point mutant ES114 DarcA::erm; arcA (VF_2120) deleted and replaced ES114 DarcA; arcA (VF_2120) gene deletion ES114 ainS::cat; C8 synthase (VF_1037) mutant ES114 DluxCDABEG ES114 DarcA::erm DluxCDABEG ES114 ainS::erm litR::kan ES114 luxI point mutant DarcA allele; oriVR6Kc, oriT, CmR DarcA::erm allele; oriVR6Kc, oriT, CmR, ErmR lacIq and IPTG-inducible promoter, oriVR6Kc, oriVpES213, oriT, KnR promoterless lacZ, oriVR6Kc, oriVpES213, oriT, CmR pEVS122 with luxI upstream sequence; oriVR6Kc, oriT, ErmR DluxI allele; oriVR6Kc, oriVColE1, oriT, ErmR, KnR DluxICDABEG allele; oriVR6Kc, oriVColE1, oriT, EmR, KnR DarcA allele (pAJ4) fused to pBluescript; oriVR6Kc, oriVColE1, oriT, CmR, AmpR pAKD601B-arcA (IPTG-inducible arcA), oriVR6Kc, oriVpES213, oriT, KnR oriVColE1, AmpR conjugative helper, oriVR6Kc, oriT, KnR oriVR6Kc, oriT, ErmR pCR-BluntII-TOPO with luxI downstream sequence, oriVColE1, KnR pCR-BluntII-TOPO with luxG downstream sequence, oriVColE1, KnR DarcA::erm allele (pAJ7) fused to pBluescript; oriVR6Kc, oriVColE1, oriT, CmR, ErmR, AmpR pAKD702 containing the luxI promoter region, oriVR6Kc, oriVpES213, oriT, CmR gfp, oriVR6Kc, oriVpES213, oriT, KnR rfp, oriVR6Kc, oriVpES213, oriT, CmR ATGAGCTCTAACCAGTTAGTTAGGTACCG TATCTAGAAAAGTCAGATAGAAGAGATTCTTA CGGCTAGCCCATGCAACCTCTCTTATTTTACATGATC ACCTGCAGGTGCGGTGAAGTTATTGAGCACAACATG [57] [56] [58] [67] [47] this study this study this study this study [52] [52] [63] [61] this study [50] [63] [52] [52] [81] [82] this study this study this study this study this study Stratagene [60] [56] [61] [61] this study [82] [59] [59] this study this study this study this study Table 1. Strains, plasmids, and oligonucleotides used in this work. Positive Feedback in V. fischeri lux Regulation Relevant characteristicsa Source or Reference aKnR, Kanamycin resistance; CmR and cat, Chloramphenicol resistance; ErmR and erm, Erythromycin resistance; AmpR, Ampicillin resistance; NxR, Nalidixic acid resistance; SpR, Spectinomycin resistance; CbR, Carbenicillin resistance. Plasmid replication origins are designated oriV with a subscript indicating the source, and oriT indicates the RP4 origin of transfer. bOligonucleotides are in the 59 to 39 orientation with introduced restriction sites underlined. doi:10.1371/journal.pone.0049590.t001 PLOS ONE | www.plosone.org 3 November 2012 | Volume 7 | Issue 11 | e49590