Plasmids

Plasmids

Here you can find a list of Bacillus subtilis plasmids:

Fusion to reporter genes

  • lacZ-fusions (chloramphenicol resistence)
  • [wiki|pAC6]: transcriptional lacZ-fusions (cat resistence)
  • [wiki|pAC7]: translational lacZ-fusions (kanamycin resistence)
  •  pBaSysBioII: gfp transcriptional fusions for high-throughput analysis of gene expression[PubMed|20150235]
  •  a suite of vectors for ectopic insertion of GFP, CFP and IYFP transcriptional fusions in single copy at the amyE and bglS loci
  •  List of GFP variants useful for various fusions (IPTG inducible Phyperspank with amyE integration) [PubMed|23956387]


Labelling of a protein with a flouresent tag

  • [wiki|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!
  • [wiki|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
  • [wiki|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!!!
  • [wiki|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|24874808]


Labelling of a protein with a triple FLAG tag

  • [wiki|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
  • [wiki|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
  • [wiki|pGP1370]: replicative plasmid for the fusion of 3x FLAG tag to the C-terminus of a protein, high expression due to the strong [gene|40C1E81BAB04BD1CC98FC57DF25D27DBDFEB5A59|degQ](Hy) promoter from [wiki|pBQ200]


Expression/ purification in B. subtilis

  • [wiki|pBP26]: expression of YFP in B. subtilis and in E. coli 
  • [wiki|pBP27]: expression of CFP in B. subtilis and in E. coli 
  • [wiki|pBQ200]: overexpression of proteins in B. subtilis under control of a strong degQ36 promoter
  • [wiki|pGP380]: expression of proteins in B. subtilis allows fusion to a Strep-tag at the N-terminus of the protein
  • [wiki|pGP382]: expression of proteins in B. subtilis allows fusion to a Strep-tag at the C-terminus of the protein
  • [wiki|pGP886]: integration vector (integrates in [gene|E77DD93E3CDCA9B81106E6AB0F18513360AAE9DB|xkdE]); allowing the expression of genes under the control of the xylose-inducible PxylA promoter in B. subtilis
  • [wiki|pGP888]: integration vector (integrates in [gene|DBBA549C2D71CF628E92AC7B659EA276E227D742|ganA]); allowing the expression of genes under the control of the xylose-inducible PxylA promoter in B. subtilis
  • [wiki|pGP1459]: integration vector (integrates in [gene|DBBA549C2D71CF628E92AC7B659EA276E227D742|ganA]) analogous to [wiki|pGP380]; expression of proteins in B. subtilis allows fusion to a Strep-tag at the N-terminus of the protein
  • [wiki|pGP1460]: integration vector (integrates in [gene|DBBA549C2D71CF628E92AC7B659EA276E227D742|ganA]) analogous to [wiki|pGP382]; expression of proteins in B. subtilis allows fusion to a Strep-tag at the C-terminus of the protein
  • [wiki|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 of E. coli

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

Construction of (markerless) deletion

  • [wiki|pMAD]: construction of markerless KOs in Gram-positive bacteria

Toolboxes for the work with B. subtilis

  • the biobrick box [PubMed|24295448]
  • a part toolbox to tune genetic expression [PubMed|27402159]



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Time of last update: 2023-02-06 15:48:28

Author of last update: Jstuelk