TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant characteristics a | Source or reference |
|---|---|---|
| Strains | ||
| B. subtilis | ||
| JH642 | trpC2 pheA1 | Laboratory stock |
| NM57 | JH642 lipM:: Kmr | This study |
| NM08 | NM57 amyE::Pxyl-lipM | This study |
| NM11 | NM57 amyE::Pxyl-lipB | This study |
| NM51 | JH642 lipL:: Spr | This study |
| NM09 | JH642 amyE::Pxyl-lipB | This study |
| NM10 | JH642 amyE::Pxyl-lipL | This study |
| NM12 | NM51 amyE::Pxyl-lipB | This study |
| NM13 | NM51 amyE::Pxyl-lipL | This study |
| CM28 | NM57 lipL:: Spr | This study |
| NM14 | CM28 amyE::Pxyl-lipB | This study |
| NM60 | JH642 lplJ::Spr | This study |
| NM65 | NM60 lipM:: Kmr | This study |
| NM67 | NM60 lipL:: Spr | This study |
| NM68 | NM65 lipL:: Cmr | This study |
| E. coli | ||
| DH5α | supE44 thi-1 ΔlacU169(f80lacZΔM15) endA1 recA1 hsdR17 gyrA96 relA1 trp6 cysT329 | Laboratory stock |
| MG1655 | rph-1 | CGSCb |
| EMM99 | E. coli BL21(DE3) / pEM88 | (Martinez et al., 2010) |
| TM131 | rpsL8 lipA::Tn1000 lplA::Tn10 | (Morris et al., 1994) |
| TM136 | rpsL8 lipB::Tn1000 lplA::Tn10 | (Morris et al., 1994) |
| JK1 | rpsL8 | (Morris et al., 1994) |
| MFH120 | JC7623 lacZΔM15 φ(fabA-lacZ)1(Hyb)cat fadBA poxB::pMFH23 | (Henry & Cronan, 1992) |
| QC031 | rpsL8 lipB::Tn1000 lplA::Tn10/ pQC003 | This study |
| QC032 | rpsL8 lipB::Tn1000 lplA::Tn10/ pQC004 | (Christensen & Cronan, 2010) |
| QC035 | rpsL8 / pBAD322G | (Christensen & Cronan, 2009) |
| QC038 | rpsL8 lipA::Tn1000 lplA::Tn10/ pBAD322G | (Christensen & Cronan, 2009) |
| QC057 | rpsL8 lipB::Tn1000 lplA::Tn10/ pBAD322G | (Christensen & Cronan, 2010) |
| QC080 | rpsL8 lipB::Tn1000 lplA::Tn10/ pQC032 | This study |
| QC081 | rpsL8 lipA::Tn1000 lplA::Tn10 / pQC032 | This study |
| QC086 | rpsL8 lipB::Tn1000 lplA::Tn10/ pQC003 | This study |
| QC087 | rpsL8 lipB::Tn1000 lplA::Tn10/ pQC004 | (Christensen & Cronan, 2010) |
| QC134 | rph-1 IN(rrnD-rrnE)1 φ(fabA-lacZ)1(Hyb)cat | (Christensen & Cronan, 2010) |
| QC142 | rph-1 IN(rrnD-rrnE)1 φ(fabA-lacZ)1(Hyb)cat / pCY598, pQC015 | (Christensen & Cronan, 2010) |
| QC143 | rph-1 IN(rrnD-rrnE)1 φ(fabA-lacZ)1(Hyb)cat / pCY598,pQC033 | This study |
| QC161 | rph-1 ΔlplA::FRT ΔlipB::FRT / pQC057, pTARA | (Christensen & Cronan, 2010) |
| QC168 | rpsL lipB::Kn lplA::Tc φ(fabA- lacZ)1(Hyb)cat | This study |
| QC175 | rpsL lipB::Kn lplA::Tc φ(fabA- lacZ)1(Hyb)cat / pBAD322G | This study |
| QC176 | rpsL lipB::Kn lplA::Tc φ(fabA- lacZ)1(Hyb)cat / pQC003 | This study |
| QC177 | rpsL lipB::Kn lplA::Tc φ(fabA- lacZ)1(Hyb)cat / pQC004 | This study |
| QC178 | rpsL (SmR) lipB::Kn lplA::Tc φ(fabA-lacZ)1(Hyb)cat / pQC032 | This study |
| Plasmids | ||
| pGES40 | pBluescript (Stratagene) containing the xylR gene and PxylA promoter | (Schujman, unpublished) |
| pJM116 | Integrative vector to construct transcriptional fusions to lacZ; integrates at the amyE locus of B. subtilis; Cmr | {Dartois, 1996 #3} |
| pJM105A | Integrational vector; Cmr | (Schujman et al., 2001, Perego, 1993) |
| pJM114 | Integrational vector; Kmr | (Schujman et al., 2001, Perego, 1993) |
| pJM134 | Integrational vector; Spr | (Schujman et al., 2001, Perego, 1993) |
| pBAD322G | arabinose inducible expression vector | (Cronan, 2006) |
| pTARA | pACYC origin, arabinose inducible T7 polymerase | (Wycuff & Matthews, 2000) |
| pCY598 | RSF origin, arabinose inducible T7 polymerase | (Cronan, 2003) |
| pET101/D-TOPO | T7 promoter expression vector | Invitrogen |
| pCR2.1 | TOPO TA cloning vector | Invitrogen |
| pNM03 | pJM134 containing lipL interrupted with an spectinomycin cassette | This study |
| pNM07 | pJM114 containing lipM interrupted with a kanamycin cassette | This study |
| pNM48 | pJM134 containing lplJ interrupted with an spectinomycin cassette | This study |
| pNM47 | pJM114 containing lipL interrupted with a kanamycin cassette | This study |
| pNM53 | pJM105A containing lipL interrupted with a chloramphenicol cassette | This study |
| pNM57 | Contains xylR PxylA-lipM into BamHI site of pJM116 | This study |
| pNM58 | Contains xylR PxylA-lipL into BamHI site of pJM116 | This study |
| pNM59 | Contains xylR PxylA-lipB into BamHI site of pJM116 | This study |
| pSJ120 | E. coli LipB expression vector | (Jordan & Cronan, 2003) |
| pQC003 | LplJ expression vector | This study |
| pQC004 | LipM expression vector | (Christensen & Cronan, 2010) |
| pQC015 | N-terminal hexahistidine LipM expression vector | (Christensen & Cronan, 2010) |
| pQC032 | LipL expression vector | This study |
| pQC033 | N-terminal hexahistidine lipL expression vector | This study |
a
Cmr, Gmr Kmr, Spr, Ampr denote resistance to chloramphenicol, gentamicin, kanamycin, spectinomycin and ampicillin, respectively.
b
CGSC denotes the Coli Genetic Stock Center.