Table 2.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant genotype/phenotype1 | Reference |
|---|---|---|
| Escherichia coli | ||
| BL21(DE3) strains | ||
| KA729 | pET22b(+)-Eflit (from E. faecalis ATCC 19433) | This study |
| KA736 | lpp::Cmr + pET22b(+)-Eflit | This study |
| KA801 | lpp::Cmr lnt::Specr chiQ::Aprr + pET22b(+)-Eflit | This study |
| KA818 | lpp::Cmr lnt::Specr chiQ::Aprr + pET22b(+)-Eflit + pKA810 | This study |
| KA893 | lpp::Cmr | This study |
| E. coli | ||
| BW25113 strains | ||
| TXM327 | lpp::Cmr | 8 |
| KA349 | lpp::Cmr ybeX-(Kanr-rrnB TT-araC-PBAD)-lnt | 11 |
| TXM541 | gut::Kanr-rrnB TT-araC-PBAd-lnt, lnt::Specr, chiQ::Aprr | 8 |
| KA775 | TXM327 lnt::Specr + pCT763 (spontaneous suppressor mutant) | Laboratory stock |
| KA827 | phoA::Kanr | Keio collection |
| KA845 | E. coli BW25113 fadR::Trimr | This study |
| SS851 | KA827 + pET22b(+)-Eflit; Carbr | This study |
| SS904 | KA827 + pET22b(+)-lacZ; Carbr | This study |
| SS915 | KA827 + pET22b(+)-EflitK212-(+9)lacZ fusion; Carbr | This study |
| SS916 | KA827 + pET22b(+)-EflitK212-(+13)phoA fusion; Carbr | This study |
| SS932 | KA827 + pET22b(+)-EflitD173-(+9)lacZ fusion; Carbr | This study |
| SS935 | KA827 + pET22b(+)-EflitR129-(+9)lacZ fusion; Carbr | This study |
| SS936 | KA827 + pET22b(+)-EflitR129-(+13)phoA fusion; Carbr | This study |
| SS940 | KA827 + pET22b(+)-EflitD173-(+13)phoA fusion; Carbr | This study |
| SS950 | KA827 + pET22b(+)-EflitD52-(+13)phoA fusion; Carbr | This study |
| SS951 | KA827 + pET22b(+)-EflitD52-(+9)lacZ fusion; Carbr | This study |
| SS964 | KA827 + pET22b(+)-phoA; Carbr | This study |
| TXM1014 | KA845 lpp::Kanr flanked by FRT sites | This study |
| TXM1015 | TXM1014 lpp::FRT | This study |
| TXM1018 | TXM1015 fadE::Tetr | This study |
| TXM1O29 | TXM1018 + pTXM1O26 | This study |
| TXM1036 | TXM1O29 lnt::Sptr | This study |
| TXM1058 | TXM1036 fabH::Cmr flanked by FRT sites | This study |
| TXM1067 | TXM1058 fabH::FRT | This study |
| TXM1111 | TXM1067 + pTM1100 | This study |
| Plasmids | ||
| pCL25 | Cloning vector with RepA origin | 39 |
| pSEVA434 | General cloning vector with pBBR1 origin | 71 |
| pKA522 | pCL25(ori) lpp(K58A)-Strep-tag Kanr | 8 |
| pET22b(+) | Expression vector with T7 promoter lacI; Carbr | Novagen |
| pET22b(+)-Eflit | pET22b(+) Eflit (from E. faecalis ATCC 19433), 212 amino acid ORF | This study |
| pET22b(+)-Eflit strep | pET22b(+) Eflit strep tag (from E. faecalis ATCC 19433) | This study |
| pET22b(+)-Eflit strep | pET22b(+) Eflit strep tag (from E. faecalis ATCC 19433) lacking all cysteine residues | This study |
| C16A C100A C187A | ||
| pLI50-Ppen Gfpmut2 | Mid-copy plasmid number with Ppen promoter controlling insert expression; Carbr | 72 |
| pLI50-Eflit strep | pLI50-Ppen lit lacking all endogenous cysteine residues | This study |
| C16A C100A C187A | ||
| pCT763 | pKA522 PA3286 (from Pseudomonas aeruginosa) | This study |
| pKA810 | pCL25(ori) Tmpr-lpp(K58A)-Strep-tag | This study |
| pTXM1O26 | pBBR1(ori) PKan-lolCDE-PA3286 Kanr | This study |
| pTM1100 | pCL25(ori) lpp(K58A)-Strep-tag-fadD(V451A) Cmr | This study |
1Resistance phenotypes: Aprr, apramycin; Carbr, carbenicillin; Cmr, chloramphenicol; Kanr, kanamycin; Sptr, spectinomycin; Tetr, tetracycline; and Tmpr, trimethoprim.