Bacterial strains and plasmids used in this study
| Strain | Genotype | Derivation | Antibiotic resistance | Source | ||||
|---|---|---|---|---|---|---|---|---|
| Bacillus anthracis Sterne 7702 derivative strains | ||||||||
| B. anthracis Sterne 7702 | Parent | None | Pasteur Institute Cataldi et al. (1990) | |||||
| B. anthracis Sterne 7702 SR1 | Streptomycin resistant | Streptomycin-resistant isolate of B. anthracis Sterne 7702 | Strep | This study | ||||
| ORB7863 | ICEBs1::Pspank(hy)-spxA1DD | ICEBs1::pCSZ35 (Pspank(hy)-spxA1DD) × B. anthracis Sterne 7702 SR1 | Spec, Strep | This study | ||||
| ORB7864 | ICEBs1::Pspank(hy)-spxA2DD | ICEBs1::pCSZ36 (Pspank(hy)-spxA2DD) × B. anthracis Sterne 7702 SR1 | Spec, Strep | This study | ||||
| ORB8092 | ICEBs1 | pJMA402 (ICEBs1) × B. anthracis Sterne 7702 SR1 | Spec, Strep | This study | ||||
| ORB8115 | ΔspxA2 | pSB3 (ΔspxA2) × B. anthracis Sterne 7702 SR1 | Strep | This study | ||||
| ORB8170 | ΔspxA1 | pSB2 (ΔspxA1) × B. anthracis Sterne 7702 | None | This study | ||||
| ORB8285 | ΔspxA1 ΔspxA2 | pSB3 (ΔspxA2) × ORB8170 | None | This study | ||||
| ORB8390 | ΔspxA2 ICEBs1::Pspank(hy)-spxA1DD | ICEBs1::pCSZ35 (Pspank(hy)-spxA1DD) × ORB8115 | Spec, Strep | This study | ||||
| ORB8391 | ΔspxA2 ICEBs1::Pspank(hy)-spxA2DD | ICEBs1::pCSZ36 (Pspank(hy)-spxA2DD) × ORB8115 | Spec, Strep | This study | ||||
| ORB8398 | ΔspxA1 | Streptomycin-resistant isolate of ORB8170 | Strep | This study | ||||
| ORB8404 | ΔspxA1 ICEBs1::Pspank(hy)-spxA1DD | ICEBs1::pCSZ35 (Pspank(hy)-spxA1DD) × ORB8170 | Spec, Strep | This study | ||||
| ORB8405 | ΔspxA1 ICEBs1::Pspank(hy)-spxA2DD | ICEBs1::pCSZ36 (Pspank(hy)-spxA2DD) × ORB8170 | Spec, Strep | This study | ||||
| ORB8438 | ΔspxA2 | pSB3 (ΔspxA2) × B. anthracis Sterne 7702 | None | This study | ||||
| ORB8481 | ΔspxA1 ΔspxA2 | Streptomycin-resistant isolate of ORB8285 | Strep | This study | ||||
| ORB8485 | ΔspxA1 ΔspxA2 ICEBs1::Pspank(hy)-spxA1DD | ICEBs1::pCSZ35 (Pspank(hy)-spxA1DD) × ORB8481 | Spec, Strep | This study | ||||
| ORB8486 | ΔspxA1 ΔspxA2 ICEBs1::Pspank(hy)-spxA2DD | ICEBs1::pCSZ36 (Pspank(hy)-spxA2DD) × ORB8481 | Spec, Strep | This study | ||||
| Bacillus subtilis JH642 (trpC2 pheA1) derivative strains | ||||||||
| JH642 | Parent | None | J. Hoch | |||||
| JMA475 | cgcD::Pspank(hy)-rapI kan458 | Kan | A. Grossman Auchtung et al. (2005) | |||||
| ORB3834 | Δspx::neo | Δspx::neo × JH642 | Neo | Nakano et al. (2001) | ||||
| ORB7262 | Δspx::neo thrC::PBA5387-lacZ | thrC::pDYR9 (PBA5387-lacZ) × ORB8384 | Neo, Erm | This study | ||||
| ORB7854 | ICEBs1::Pspank(hy)-spxA1DD | ICEBs1::pCSZ35 (Pspank(hy)-spxA1DD) × JH642 | Spec | This study | ||||
| ORB7860 | ICEBs1::Pspank(hy)-spxA2DD | ICEBs1::pCSZ36 (Pspank(hy)-spxA2DD) × JH642 | Spec | This study | ||||
| ORB7861 | ICEBs1::Pspank(hy)-spxA1DD cgcD::Pspankhy-rapI kan458 | JMA475 genomic DNA × ORB7854 | Spec, Kan | This study | ||||
| ORB7862 | ICEBs1::Pspank(hy)-spxA2DD cgcD::Pspank(hy)-rapI kan458 | JMA475 genomic DNA × ORB7860 | Spec, Kan | This study | ||||
| ORB7871 | Δspx thrC::PBA5387-lacZ amyE::Pspank(hy)-spxA1DD | amyE::pMMN818 (Pspank(hy)-spxA1DD) × ORB7262 | Neo, Erm, Spec | This study | ||||
| ORB7872 | Δspx thrC::PBA5387-lacZ amyE::Pspank(hy)-spxA2DD | amyE::pMMN819 (Pspank(hy)-spxA2DD) × ORB7262 | Neo, Erm, Spec | This study | ||||
| ORB8343 | Δspx::neo thrC::PBA0847-lacZ | thrC::PBA0847-lacZ × ORB3834 | Neo, Erm | This study | ||||
| ORB8344 | Δspx::neo thrC::PBA1119-lacZ | thrC::PBA1119-lacZ × ORB3834 | Neo, Erm | This study | ||||
| ORB8354 | Δspx::neo thrC::PBA1951-lacZ | thrC::PBA1951-lacZ × ORB3834 | Neo, Erm | This study | ||||
| ORB8356 | Δspx::neo thrC::PBA5387-lacZ | thrC::PBA5387-lacZ × ORB3834 | Neo, Erm | This study | ||||
| ORB8359 | Δspx::neo thrC::PBA0847-lacZ amyE::Pspank(hy)-spxA1DD | amyE:: Pspank(hy)-spxA1DD × ORB8343 | Neo, Erm, Spec | This study | ||||
| ORB8360 | Δspx::neo thrC::PBA0847-lacZ amyE:: Pspank(hy)-spxA2DD | amyE:: Pspank(hy)-spxA2DD × ORB8343 | Neo, Erm, Spec | This study | ||||
| ORB8361 | Δspx::neo thrC::PBA1119-lacZ amyE:: Pspank(hy)-spxA1DD | amyE:: Pspank(hy)-spxA1DD × ORB8344 | Neo, Erm, Spec | This study | ||||
| ORB8362 | Δspx::neo thrC::PBA1119-lacZ amyE:: Pspank(hy)-spxA2DD | amyE:: Pspank(hy)-spxA2DD × ORB8344 | Neo, Erm, Spec | This study | ||||
| ORB8363 | Δspx::neo thrC::PBA1951-lacZ amyE:: Pspank(hy)-spxA1DD | amyE:: Pspank(hy)-spxA1DD × ORB8354 | Neo, Erm, Spec | This study | ||||
| ORB8364 | Δspx::neo thrC::PBA1951-lacZ amyE:: Pspank(hy)-spxA2DD | amyE:: Pspank(hy)-spxA2DD × ORB8354 | Neo, Erm, Spec | This study | ||||
| ORB8367 | Δspx::neo thrC::PBA5387-lacZ amyE:: Pspank(hy)-spxA1DD | amyE:: Pspank(hy)-spxA1DD × ORB8356 | Neo, Erm, Spec | This study | ||||
| ORB8368 | Δspx::neo thrC::PBA5387-lacZ amyE:: Pspank(hy)-spxA2DD | amyE:: Pspank(hy)-spxA2DD × ORB8356 | Neo, Erm, Spec | This study | ||||
| ORB8373 | Δspx::neo thrC::PBA1200-lacZ | thrC::PBA1200-lacZ × ORB3834 | Neo, Erm | This study | ||||
| ORB8380 | Δspx::neo thrC::PspxA1-lacZ amyE:: Pspank(hy)-spxA1DD | amyE:: Pspank(hy)-spxA1DD × ORB8373 | Neo, Erm, Spec | This study | ||||
| ORB8381 | Δspx::neo thrC::PBA1200-lacZ amyE:: Pspank(hy)-spxA2DD | amyE:: Pspank(hy)-spxA2DD × ORB8373 | Neo, Erm, Spec | This study | ||||
| ORB8389 | thrC::PBA3868-lacZ | thrC::PBA3868-lacZ × ORB3834 | Neo, Erm | This study | ||||
| ORB8396 | thrC::PBA3868-lacZ amyE:: Pspank(hy)-spxA2DD | amyE:: Pspank(hy)-spxA2DD × ORB8389 | Neo, Erm, Spec | This study | ||||
| ORB8397 | thrC::PBA3868-lacZ amyE:: Pspank(hy)-spxA1DD | amyE:: Pspank(hy)-spxA1DD × ORB8389 | Neo, Erm, Spec | This study | ||||
| Plasmid | Genotype | Selection | ||||||
| pCSZ35 | pJMA402::Pspank(hy)-spxA1DD | Spec | ||||||
| pCSZ36 | pJMA402::Pspank(hy)-spxA2DD | Spec | ||||||
| pSB2 | pRP1028::ΔspxA1 | Spec | ||||||
| pSB3 | pRP1028::ΔspxA2 | Spec | ||||||
| pSB10 | pPROEX-1::spxA2 | Amp | ||||||
| pMMN818 | pDR111::Pspank(hy)-spxA1DD | Spec | ||||||
| pMMN819 | pDR111::Pspank(hy)-spxA2DD | Spec | ||||||
| pDG793 | thrC integration vector with promoter-less lacZ reporter | Erm Guerout-Fleury et al. (1996) | ||||||
| pSS1827 | Conjugation helper strain | Amp | ||||||
| pSS4332 | Harbors I-SceI gene and cyan fluorescent protein reporter construct | Kan | ||||||
| pRP1028 | Cloning vector for conjugation | Spec | ||||||
| pJMA402 | Cloning vector for ICEBs1 conjugation | Spec | ||||||
| pDR111 | amyE integration vector with Pspank(hy) promoter | Spec | ||||||
| pPROEX-1 | E. coli expression vector | Amp | ||||||
| pDRY9 | thrC integration plasmid carrying B. subtilis trxB (−115 to +47)-lacZ | Erm | ||||||
Spec, spectinomycin; Strep, streptomycin; Kan, kanamycin; Amp, ampicillin; Erm, erythromycin; Neo, neomycin.