A facile and robust T7-promoter-based high-expression of heterologous proteins in Bacillus subtilis

Bioresources and Bioprocessing

Table 2 Bacterial strains and plasmids

Strain or plasmid	Characteristics	Source
Bacillus subtilis
SCK6	Erm^R, his nprR2 nprE18 △aprA3 △eglS102 △bglT bglSRV, lacA::P_xylA-comK	Lab stock
SCK8	Erm^R, SCK6 derivate, △upp	This work
SCK9	Erm^R, SCK8 derivate, △upp△spoIIAC	This work
SCK10	Erm^R, SCK9 derivate, △upp△spoIIAC△srfAC	This work
SCK22	Erm^R, SCK10 derivate, △upp△spoIIAC△srfAC, amyE::P₄₃-T7RNAP	This work
Plasmids
pSS	Amp^R, Cm^R, modular vector carrying upp-cassette	Lab stock
pHT01	Amp^R, Cm^R, E. coli-B. subtilis shuttle vector	Lab stock
pDG1730	Spc^R, integration vector contains a spectinomycin resistance gene sandwiched between amyE-front and amyE-back	Lab stock
pDG1730-T7RNAP	Spc^R, pDG1730 derivate, used to integrate genes on the genome, with T7RNAP expression cassette (spc-upp-P₄₃-T7RNAP) and amyE gene upstream and downstream homology arms	This work
pHT7	Amp^R, Cm^R, pHT01 derivate, with T7-lac promoter	This work
pHT7-GFP	Amp^R, Cm^R, pHT7 derivate, with GFP cloned	This work
pHT7-αGP	Amp^R, Cm^R, pHT7 derivate, with αGP cloned	This work
pHT7-IMP	Amp^R, Cm^R, pHT7 derivate, with IMP cloned	This work
pHT7-PGM	Amp^R, Cm^R, pHT7 derivate, with PGM cloned	This work
pHT7-4GT	Amp^R, Cm^R, pHT7 derivate, with 4GT cloned	This work