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Table 1 Plasmids and strains

From: Development of a novel strategy for robust synthetic bacterial promoters based on a stepwise evolution targeting the spacer region of the core promoter in Bacillus subtilis

Plasmids and strains Relevant characteristics References or source
Plasmids
 pAX01 B. subtilis integration vector, xylr-Pxyl cassette, Ampr, Ermr Lab stock
 pAX-comK Derived from pAX01, comK inserted This study
 pBP43GFP E. coli-B. subtilis shuttle vector, P43 promoter, gfp, Ampr, Kanr Lab stock
 pBSG03 E. coli-B. subtilis shuttle vector, PsrfA promoter, gfp, Ampr, Kanr Lab stock
 pBS-gusA gusA expressed by wild type PsrfA This study
 pBBH4-gusA gusA expressed by mutant promoter PBH4 This study
 pBS-NK Secretion expression of nattokinase by wild type PsrfA and WapA signal peptide This study
 pBBH4-NK Secretion expression of nattokinase by wild type PBH4 and WapA signal peptide This study
 pBPylbP-GFP Derived from pBSG03, PsrfA::PylbP-RBS, gfp, Ampr, Kanr This study
 pBPxylA-GFP Derived from pBSG03, PsrfA::PxylA, gfp, Ampr, Kanr This study
Strains
 E.coli JM109 recA1, supE44 endA1 hsdR17 (r−k,m+ k) gyrA96 relA1 thi (lac-proAB) F’[traD36 proAB+ lacIq lacZ ΔM15] Lab stock
 B. subtilis 168 trpC2 Lab stock
 B. subtilis comK Derived from B. subtilis 168, lacA::PxylA-comK, Ermr This study
 B. subtilis 3NA spo0A3 Lab stock
 BSGus B. subtilis comK harboring pBS-gusA This study
 BSBHGus B. subtilis comK harboring pBBH4-gusA This study
 BSNK B. subtilis 3NA harboring pBS-NK This study
 BSBHNK B. subtilis 3NA harboring pBBH4-NK This study