TABLE 1.
Plasmids and strains
| Plasmid or strain | Genotypea | Source or reference |
|---|---|---|
| Plasmids | ||
| pDSW204 | IPTG regulated promoter, pBR322 origin; Amp | 40 |
| pDSW207 | pDSW204-gfp-MCS (fusion vector) | 40 |
| pCR27 | pDSW207 containing myc3 | 30 |
| pMDG1 | pNG162 containing zapA-malFcyto-ftsB | M. D. Gonzalez |
| pNG162 | IPTG regulated promoter (pDSW204), pSC101 origin; Spc | 12 |
| pKDW38 | pNG162 containing zapA-malFcyto-CIBTIB-myc3; Spc | This study |
| pKDW39 | pNG162 containing zapA-malFcyto-CIBTIBαIBβIBγIB-myc3; Spc | This study |
| pKDW40 | pNG162 containing zapA-malFcyto-CIBTIBαIB-myc3; Spc | This study |
| pKDW42 | pNG162 containing zapA-malFcyto-CIBTIBβIBγIB-myc3; Spc | This study |
| pKDW48 | pNG162 containing zapA-malFcyto-CIBTIBαIBβIB-myc3; Spc | This study |
| pKDW49 | pNG162 containing zapA-malFcyto-CTRTTRαIBβIBγIB-myc3; Spc | This study |
| pSG1729 | B. subtilis amyE integration vector; Pxylbla spc | 21 |
| pKDW44 | pSG1729 containing E. coli FtsQ; Pxylbla spc | This study |
| Strains | ||
| E. coli | ||
| JOE309 | MC4100 ara+ | 3 |
| CR362 | JOE309 Δ(λattL-lom)::bla lacIq/pDSW204-gfp-pbpB-myc3 | 30 |
| CR363 | JOE309 Δ(λattL-lom)::bla lacIq/pDSW204-gfp-pbpB | 30 |
| B. subtilis | ||
| SU5 (or 168) | trpC2 | Laboratory collection |
| RSA8 | trpC2 divIB::cat::ermC | 31 |
| RSA9 | trpC2 divIB::cat::ermC amyE::divIB cat | 31 |
| RSA24 | trpC2 divIB::cat::ermC amyE::divIB(E122A) cat | This study |
| RSA16 | trpC2 divIB::cat::ermC amyE::divIB(L139A) cat | This study |
| RSA18 | trpC2 divIB::cat::ermC amyE::divIB(N141A) cat | This study |
| RSA26 | trpC2 divIB::cat::ermC amyE::divIB(E187A) cat | This study |
| RSA19 | trpC2 divIB::cat::ermC amyE::divIB(Y203A) cat | This study |
| RSA21 | trpC2 divIB::cat::ermC amyE::divIB(N205A) cat | This study |
| RSA22 | trpC2 divIB::cat::ermC amyE::divIB(D206A) cat | This study |
| RSA30 | trpC2 divIB::cat::ermC amyE::divIB(Y224A) cat | 31 |
| RSA31 | trpC2 divIB::cat::ermC amyE::divIB(P225A) cat | 31 |
| RSA28 | trpC2 divIB::cat::ermC amyE::divIB(Y246A) cat | This study |
a
In the plasmid notation, the subscripts IB and TR indicate that the domains are from B. subtilis DivIB and E. coli TolR, respectively, and C and T refer to the cytoplasmic and transmembrane domains, respectively. The three extracytoplasmic domains of DivIB are referred to as α, β, and γ (31). MCS, multiple cloning site; cyto, cytoplasmic region.