Table 1.
Strains and plasmids used in this study
| Strain | Description | Source |
|---|---|---|
| B. subtilis | ||
| 168 | trpC2 | BGSCa |
| 168 Cel8A | trpC2 thrC::pCel8A, ery | This work |
| 168 Cel8A-LysM | trpC2 thrC::pCel8A-LysM, ery | This work |
| 168 Cel8A-[L]-LysM | trpC2 thrC::pCel8A-[L]-LysM, ery | This work |
| BRB07 | trpC2, nprB, aprE, epr, bpr, nprE, mpr, vpr | Cobra Biologics (Pohl, Bhavsar et al. 2013) |
| BRB07 Cel8A |
trpC2, nprB, aprE, epr, bpr, nprE, mpr, vpr thrC::pCel8A, ery |
This work |
| BRB07 Cel8A-LysM |
trpC2, nprB, aprE, epr, bpr, nprE, mpr, vpr thrC::pCel8A-LysM, ery |
This work |
| BRB07 Cel8A-[L]-LysM |
trpC2, nprB, aprE, epr, bpr, nprE, mpr, vpr thrC::pCel8A-[L]-LysM, ery |
This work |
| BRB08 | trpC2, nprB, aprE, epr, bpr, nprE, mpr, vpr, wprA | Cobra Biologics (Pohl, Bhavsar et al. 2013) |
| BRB08 Cel8A |
trpC2, nprB, aprE, epr, bpr, nprE, mpr, vpr, wprA thrC::pCel8A, ery |
This work |
| BRB08 Cel8A-LysM |
trpC2, nprB, aprE, epr, bpr, nprE, mpr, vpr, wprA thrC::pCel8A-LysM, ery |
This work |
| BRB08 Cel8A-[L]-LysM |
trpC2, nprB, aprE, epr, bpr, nprE, mpr, vpr, wprA thrC::pCel8A-[L]-LysM, ery |
This work |
| BRB14 |
trpC2, nprB, aprE, epr, bpr, nprE, mpr, vpr, wprA, htrA, htrB |
Cobra Biologics (Pohl, Bhavsar et al. 2013) |
| BRB14 Cel8A |
trpC2, nprB, aprE, epr, bpr, nprE, mpr, vpr, wprA, htrA, htrB thrC::pCel8A, ery |
This work |
| BRB14 Cel8A-LysM |
trpC2, nprB, aprE, epr, bpr, nprE, mpr, vpr, wprA, htrA, htrB thrC::pCel8A-LysM, ery |
This work |
| BRB14 Cel8A-[L]-LysM |
trpC2, nprB, aprE, epr, bpr, nprE, mpr, vpr, wprA, htrA, htrB thrC::pCel8A-[L]-LysM, ery |
This work |
| WB800 |
trpC2, nprB::bsr, aprE, epr, bpr, nprE, mpr::ble, vpr, wprA::hyg (cat) |
(Wu, Yeung et al. 2002) |
| WB800S |
trpC2, nprB::bsr, aprE, epr, bpr, nprE, mpr::ble, vpr, wprA::hyg, cat::spc |
This work |
| WB800S Cel8A |
trpC2, nprB::bsr, aprE, epr, bpr, nprE, mpr::ble, vpr, wprA::hyg, cat::spc thrC::pCel8A, ery |
This work |
| WB800S Cel8A-LysM |
trpC2, nprB::bsr, aprE, epr, bpr, nprE, mpr::ble, vpr, wprA::hyg, cat::spc thrC::pCel8A-LysM, ery |
This work |
| WB800S Cel8A-[L]-LysM |
trpC2, nprB::bsr, aprE, epr, bpr, nprE, mpr::ble, vpr, wprA::hyg, cat::spc thrC::pCel8A-[L]-LysM, ery |
This work |
| WB800S LysM-Cel8A |
trpC2, nprB::bsr, aprE, epr, bpr, nprE, mpr::ble, vpr, wprA::hyg, cat::spc thrC::pLysM-Cel8A, ery |
This work |
| WB800S LysM-[L]-Cel8A |
trpC2, nprB::bsr, aprE, epr, bpr, nprE, mpr::ble, vpr, wprA::hyg, cat::spc thrC::pLysM-[L]-Cel8A, ery |
This work |
| E. coli | ||
| BL21 (DE3) |
fhuA2 [lon] ompT gal (λ DE3) [dcm] ∆hsdS λ DE3 = λ sBamHIo ∆EcoRI-B int::(lacI::PlacUV5::T7 gene1) i21 ∆nin5 |
Novagen |
| BL21 (DE3) Cel8A |
fhuA2 [lon] ompT gal (λ DE3) [dcm] ∆hsdS λ DE3 = λ sBamHIo ∆EcoRI-B int::(lacI::PlacUV5::T7 gene1) i21 ∆nin5 pE-SUMO- Cel8A |
This work |
| Plasmid | ||
| pJL62 |
B. subtilis – E. coli shuttle vector that converts cam to scp; amp, tet |
(Ledeaux and Grossman 1995) |
| pBL113 |
B. subtilis – E. coli shuttle vector derived from pRDC19 (Arigoni, Talabot et al. 1998) that integrates into the thrC locus in the B. subtilis genome, amp, erm, IPTG inducible |
B. Lazazzera, UCLA (unpublished) |
| pCel8A | PhrC signal peptide (1–35) preceding His6 tagged Cel8A (32–394) in pBL113 |
This work |
| pCel8A-LysM | PhrC signal peptide (1–35) preceding His6 tagged Cel8A (32–394) and LysM domains (25–230) in pBL113 |
This work |
| pCel8A-[L]-LysM | PhrC signal peptide (1–35) preceding His6 tagged Cel8A (32–394), linker region from FnBPB (770–880) and LysM domains (25–230) in pBL113 |
This work |
| pLysM-Cel8A | PhrC signal peptide (1–35) preceding His6 tagged LysM domains (25–230) and Cel8A (32–394) in pBL113 |
This work |
| pLysM-[L]-Cel8A | PhrC signal peptide (1–35) preceding His6 tagged LysM domains (25–230), linker region from FnBPB (770–880) and Cel8A (32–394) in pBL113 |
This work |
| pE-SUMO |
E. coli plasmid for protein overexpression and purification, kan, IPTG inducible |
LifeSensors |
| pE-SUMO-Cel8A | Overexpression of the Cel8A (32–394) catalytic domain in pE-SUMO |
This work |
Table caption: amp, ble, bsr, cam, ery, hyg, kan, spc, and tet are antibiotic resistance markers for ampicillin, bleomycin, blasticidin S, chloramphenicol, erythromycin, hygromycin, kanamycin, spectinomycin, and tetracycline respectively.
a
BGSC, Bacillus Genetic Stock Center.
GenBank accession numbers are given in the materials and methods.