List of plasmids

Here you can find a list of Bacillus subtilis plasmids:

Fusion to reporter genes

• pAC5: translational lacZ-fusions (chloramphenicol resistence)
• pAC6: transcriptional lacZ-fusions (cat resistence)
• pAC7: translational lacZ-fusions (kanamycin resistence)
• pBaSysBioII: gfp transcriptional fusions for high-throughput analysis of gene expression PubMed
• a suite of vectors for ectopic insertion of GFP, CFP and IYFP transcriptional fusions in single copy at the amyE and bglS loci PubMed
• List of GFP variants useful for various fusions (IPTG inducible Phyperspank with amyE integration) PubMed

Labelling of a protein with a flourescent tag

• pBP43: integrative plasmid for the fusion of monomeric GFP tag to the C-terminus of a protein, keeping the control of the expression under its natural promoter, this is the preferred plasmid for constructing GFP fusions!
• pGP1871: integrative plasmid for the fusion of YFP tag to the C-terminus of a protein, keeping the control of the expression under its natural promoter
• pGP1080: integrative plasmid for the fusion of GFP tag to the C-terminus of a protein, keeping the control of the expression under its natural promoter, allows express of the downstream gene by the P(spac) promoter, ATTENTION: This plasmid encodes the dimeric GFP that tends to produce artifacts!!!
• pGP1870: integrative plasmid for the fusion of GFP tag to the C-terminus of a protein, keeping the control of the expression under its natural promoter, ATTENTION: This plasmid encodes the dimeric GFP that tends to produce artifacts!!!
• ectopic integration vectors for generating fluorescent promoter fusions in Bacillus subtilis with minimal dark noise PubMed

Labelling of a protein with a triple FLAG tag

• pGP1087: integrative plasmid for the fusion of 3x FLAG tag to the C-terminus of a protein, keeping expression under the control of the natural promoter, allowing the expression of downstream genes under the control of the IPTG-inducible Pspac promoter
• pGP1331: integrative plasmid for the fusion of 3x FLAG tag to the C-terminus of a protein, keeping expression under the control of the natural promoter
• pGP1370: replicative plasmid for the fusion of 3x FLAG tag to the C-terminus of a protein, high expression due to the strong degQ(Hy) promoter from pBQ200

Expression/ purification in B. subtilis

• pBP26: expression of YFP in B. subtilis and in E. coli
• pBP27: expression of CFP in B. subtilis and in E. coli
• pBQ200: overexpression of proteins in B. subtilis under control of a strong degQ36 promoter
• pGP380: expression of proteins in B. subtilis allows fusion to a Strep-tag at the N-terminus of the protein
• pGP382: expression of proteins in B. subtilis allows fusion to a Strep-tag at the C-terminus of the protein
• pGP886: integration vector (integrates in xkdE); allowing the expression of genes under the control of the xylose-inducible PxylA promoter in B. subtilis
• pGP888: integration vector (integrates in ganA); allowing the expression of genes under the control of the xylose-inducible PxylA promoter in B. subtilis
• pGP1459: integration vector (integrates in ganA) analogous to pGP380; expression of proteins in B. subtilis allows fusion to a Strep-tag at the N-terminus of the protein
• pGP1460: integration vector (integrates in ganA) analogous to pGP382; expression of proteins in B. subtilis allows fusion to a Strep-tag at the C-terminus of the protein
• pGP1389: integrative plasmid for the fusion of a Strep-tag to the C-terminus of a protein, keeping expression under the control of the natural promoter

Expression/ purification in E. coli

• pWH844: expression of proteins in E. coli, allows fusion to a N-terminal His-tag
• pGP570: expression of proteins in E. coli, allows fusion to a N-terminal His-tag that can be cleaved off by thrombin
• pETM-11: expression of proteins in E. coli, allows fusion to a N-terminal His-tag that can be cleaved off by TEV protease
• pGP172: expression of proteins in E. coli allows fusion to a Strep-tag at the N-terminus of the protein
• pGP574: expression of proteins in E. coli allows fusion to a Strep-tag at the C-terminus of the protein

Construction of (markerless) deletions

• pMAD: construction of markerless KOs in Gram-positive bacteria

Toolboxes for the work with B. subtilis

• the biobrick box PubMed
• a part toolbox to tune genetic expression PubMed