TABLE 1.
Bacterial strains, plasmids, and oligonucleotides
| Strain, plasmid, or oligonucleotide | Relevant characteristicsa | Source or reference |
|---|---|---|
| V. fischeri | ||
| ES114 | Wild-type isolate from Euprymna scolopes | 3 |
| EW1 | ES114 Δphr (allele exchanged from pELW1) | This study |
| EW4 | ES114 Δphr ΔluxCDABEG (EW1 with lux allele exchanged from pEVS153) | This study |
| EVS102 | ES114 ΔluxCDABEG | J. L. Bose and E. V. Stabb, unpublished data |
| JRM100 | ES114 mini-Tn7-ermR (inserted at intergenic att site) | 26 |
| E. coli | ||
| CC118λpir | Δ(ara-leu) araD ΔlacX74 galE galK phoA20 thi-1 rpsE rpoB argE(Am) recA1; lysogenized with λpir | 20 |
| DH5α | F−F80dlacZΔM15 Δ(lacZYA-argF)U169 deoR supE44 hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | 14 |
| DH5αλpir | DH5α lysogenized with λpir | 10 |
| Plasmids | ||
| pCR-Blunt II-TOPO | PCR product cloning vector; ColE1 oriV kanR | Invitrogen |
| pALZ1 | PCR product (primers JBPHOTO1 and JBPHOTO2; ES114 template) in pCR-Blunt II-TOPO | This study |
| pALZ5 | SpeI-AvrII fragment of pALZ1 containing phr and flanking sequences cloned between SpeI-AvrII sites of pEVS118 | This study |
| pALZ6 | PCR product (primers JBPHOTO3 and JBPHOTO4; pALZ5 template) SmaI digested and self-ligated | This study |
| pBluescript KS(+) | ColE1 oriV ampR | Stratagene |
| pELW1 | SpeI-digested pALZ6 ligated to SpeI-digested pBluescript | This study |
| pELW2 | PCR product (primers JBPHOTO9 and JBPHOTO10; pALZ1 template) in pCR-Blunt II-TOPO | This study |
| pELW5 | phr-containing AvrII-KpnI fragment of pELW2 cloned between SpeI-KpnI sites of pVSV105 | This study |
| pEVS104 | R6Kγ oriV tra trb oriTRP4kanR; conjugative helper | 37 |
| pEVS118 | R6Kγ oriV oriTRP4chmR | 10 |
| pEVS153 | ColE1 oriV kanR chmR oriTRP4 with ΔluxCDABEG and flanking sequence from ES114 | Bose and Stabb, unpublished |
| pVSV105 | pES213 oriV chmR oriTRP4 | 11 |
| Oligonucleotides | ||
| JBPHOTO1 | 5′ GCT CTA GAG GTC TGG CAA GGT ACT TAA TAC GGC T 3′ | This study |
| JBPHOTO2 | 5′ CCG CCT AGG TGG GTG CCG CTG ACA CAA TAA TGA 3′ | This study |
| JBPHOTO3 | 5′ TCC CCC GGG CAT AGC GTA ATC TTA GCT CTT GTG GGT AAG 3′ | This study |
| JBPHOTO4 | 5′ TCC CCC GGG TAA CCA CGC TTG AGT ATG ATT AAT ACA TC 3′ | This study |
| JBPHOTO5 | 5′ CGC ATC GCC TTA GTT GAT TCT AAG GAT TC 3′ | This study |
| JBPHOTO6 | 5′ CAC CTC AGA AGC AGG CTT TAG CTG AGC TC 3′ | This study |
| JBPHOTO7 | 5′ GCC CCA CTC TGA AAT ACA AAA TTG CTC AG 3′ | This study |
| JBPHOTO8 | 5′ GTG TGG GTT GCG GCA CGA GCC ATG 3′ | This study |
| JBPHOTO9 | 5′ CCG CCC TAG GCA AGA GCT AAG ATT ACG CTA TGG AT 3′ | This study |
| JBPHOTO10 | 5′ GGG GTA CCG GAT GCC TTG CGA TGG GGT TAT ATC 3′ | This study |
a
ampR, ampicillin resistance (bla); chmR, chloramphenicol resistance (cat); ermR, erythromycin resistance; and kanR, kanamycin resistance (aph).