Table 1.
| Strain | Genotype | Source |
|---|---|---|
| Escherichia coli: | ||
| MG1655 | rph-1 | CGSCa |
| RMK98 | MC1061 fadK::his6-cat ΔompT::FRT | this study |
| QC146 | rph-1 ΔlplA::FRT ΔlipB::FRT | (Christensen & Cronan, 2009) |
| QC152 | rph-1 ΔlplA::FRT ΔlipB::FRT/pTARA, pQC026 | this study |
| QC172 | rph-1 ΔlplA::FRT ΔlipB::FRT ΔompT::FRT | this study |
| QC191 | rph-1 ΔlplA::FRT ΔlipB::FRT ΔompT::FRT/pQC047, pTARA | this study |
| QC222 | rph-1 IN(rrnD-rrnE)1 ϕ(fabA-lacZ)1(Hyb)cat/pQC077, pCY598 | this study |
| QC223 | rph-1 IN(rrnD-rrnE)1 ϕ(fabA-lacZ)1(Hyb)cat/pQC078, pCY598 | this study |
| QC224 | rph-1 IN(rrnD-rrnE)1 ϕ(fabA-lacZ)1(Hyb)cat/pQC079, pCY598 | this study |
| QC225 | rph-1 IN(rrnD-rrnE)1 ϕ(fabA-lacZ)1(Hyb)cat/pQC080, pCY598 | this study |
| QC255 | rph-1 IN(rrnD-rrnE)1 ϕ(fabA-lacZ)1(Hyb)cat/pQC086, pCY598 | this study |
| QC256 | rph-1 IN(rrnD-rrnE)1 ϕ(fabA-lacZ)1(Hyb)cat/pQC087, pCY598 | this study |
| QC257 | rph-1 IN(rrnD-rrnE)1 ϕ(fabA-lacZ)1(Hyb)cat/pQC088, pCY598 | this study |
| QC258 | rph-1 IN(rrnD-rrnE)1 ϕ(fabA-lacZ)1(Hyb)cat/pQC089, pCY598 | this study |
| Bacillus subtilis: | ||
| JH642 | trpC2 pheA1 | Laboratory stock |
| NM20 | JH642 gcvH:: Km | This study |
| NM21 | NM20 amyE::Pspac-gcvH | This study |
| Plasmids | Relevant Properties | Source |
| pTARA | pACYC origin, arabinose inducible T7 polymerase | (Wycuff & Matthews, 2000) |
| pCY598 | RSF origin, arabinose inducible T7 polymerase | (Cronan, 2003) |
| pCR2.1 | TOPO TA cloning vector | Invitrogen |
| pSJ120 | E. coli LipB expression vector | (Jordan & Cronan, 2003) |
| pQC023 | B. subtilis E2p lipoyl domain expression vector | this study |
| pQC026 | C-terminal hexahistidine BLAP expression vector | this study |
| pQC033 | N-terminal hexahistidine LipL expression vector | (Martin et al., 2011) |
| pQC047 | C-terminal hexahistidine PdhC expression vector | this study |
| pQC077 | N-terminal hexahistidine LipL K162R mutant expression vector | this study |
| pQC078 | N-terminal hexahistidine LipL K162A mutant expression vector | this study |
| pQC079 | N-terminal hexahistidine LipL C150A mutant expression vector | this study |
| pQC080 | N-terminal hexahistidine LipL C150S mutant expression vector | this study |
| pQC086 | N-terminal hexahistidine LipL C39A mutant expression vector | this study |
| pQC087 | N-terminal hexahistidine LipL C39S mutant expression vector | this study |
| pQC088 | N-terminal hexahistidine LipL Y180A mutant expression vector | this study |
| pQC089 | N-terminal hexahistidine LipL Y180F mutant expression vector | this study |
| pGES485 | Integrational vector; Sp | (GE Schujman, unpublished) |
| pJM114 | Integrational vector; Km | (Perego, 1993) |
| pNM67 | pJM114 containing gcvH interrupted with a kanamycin cassette | This study |
| pNM70 | pGES485 containing gcvH cloned into HindIII-BamHI sites | This study |
a
CGSC, E. coli Genetic Stock Center