Strain or plasmid | Relevant characteristics | Source or reference |
---|---|---|
E. coli | ||
 Stable | F’ proA + B + lacIq ∆(lacZ)M15 zzf::Tn10 (TetR) ∆(ara-leu) 7697 araD139 fhuA ∆lacX74 galK16 galE15 e14-Φ80dlacZ∆M15 recA1 relA1 endA1 nupG rpsL (StrR) rph spoT1 ∆(mrr-hsdRMS-mcrBC) | New England Biolabs |
 C43(DE3) | F– ompT gal dcm hsdSB(r −B m −B ) (DE3) | [55] |
B. subtilis | ||
 WB800N | nprE aprE epr bpr mpr::ble nprB::bsr∆vpr, wprA::hyg pB-cat5-neo-cat3 (NeoR) | [14] |
 KO7 | ∆nprE,,∆aprE, ∆epr, ∆mpr, ∆nprB, ∆vpr, ∆bpr | Zeigler, D. R.; Bacillus Genetic Stock Center, Columbus, OH, USA |
 KO7A | KO7, ∆wprA (HygR) | This work |
Plasmids | ||
 pBE-S | Vector used with the B. subtilis Secretory Protein Expression System, KmR, AmpR | TaKaRa/Clontech |
 pET11M-senP2 | Expression plasmid for SenP2 protease | H. Besir, EMBL Protein Expression and Purification Core Facility, Heidelberg, Germany |
 pETM11-SUMO3GFP | T7 Expression vector for E. coli encoding His6-SUMO3-eGFP fusion protein, KanR | H. Besir, EMBL Protein Expression and Purification Core Facility, Heidelberg, Germany |
 pET52B-senP2 | SenP2 encoding region N-terminally fused to StrepII-tag | This work |
 pHT01 | IPTG-inducible expression vector for B. subtilis carrying Pgrac and CmR | [49] |
 pJH-scFv | Phagemid cassette vector, IPTG-inducible expression of anti-GFP ScFv-pIII fusion protein, AmpR | This work |
 pJHS01 | pHT01 cured for MluI sites | This work |
 pJHS02 | pJHS01 encoding fusion protein consisting of AmyQ signal peptide, StrepII-tag, SUMO-tag and 6xHis-tag | This work |
 pJHS03 | pJHS02 with cloned phoA | This work |
 pJHS04-07 | pJHS03 with signal peptide encoding region changed (SPaprE (04), SPyncM (05), SPyopL,(06) and SPyoaW (07)) | This work |
 pJHS08 | pJHS02 with cloned GFP-specific scFv gene | This work |
 pJHS09 | pJHS02 with SPamyQ changed for SPyoaW | This work |
 pJHS10 | pJHS09 with cloned GFP-specific scFv gene | This work |
 pJHS11 | pJHS09 with codon usage optimized SUMO-scFv fragment | This work |
 pJHS12 | pJHS09 with codon-usage optimized SUMO-cassette | This work |
 pJHS13 | pJHS02 with cloned barnase and barstar | This work |
 pJHS14 | pJHS12 with cloned barnase and barstar | This work |
 pPA4 | vector with cloned phoA | [50] |
 pQEGFP | pQE30 (Qiagen) with cloned gfp | [53] |