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Bacillus subtilis Protein Expression System

BSUB SURE VECTORS - Bacillus subtilis SURE System vectors, 10µg

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The SURE® System: Subtilin Regulated gene Expression

Bacillus subtilis is an attractive host for gram-positive bacteria as it is non-pathogenic, does not have significant bias in codon usage, and is capable of secreting functional extracellular proteins directly into the culture medium. It is used in a variety of applications, including agricultural, medical and food biotechnology and the production of recombinant proteins.

Controlled gene expression in B. subtilis is a cost-effective alternative to IPTG induced gene expression. This is achieved with the used of subtilin, a small peptide antibiotic of 32 amino acids produced by B. subtilis. Subtilin production and regulation is encoded in the chromosome by a cluster of 9 genes, which are transcribed from two promoters. Subtilin regulates/activates its own biosynthesis via a two-component regulatory system.

In the SURE system, regulatory components called SpaR (response regulator) and SpaK (membrane sensor) were isolated and placed on the chromosome of a B. subtilis host strain. One of the subtilin regulated promoters is located upstream of a multiple cloning site into which the gene of interest can be cloned (gene X). Upon addition of subtilin, the system is activated and the protein of interest is produced. It accumulates either intracellularly or is secreted into the medium.

Fig 1: Components of the SURE system

Fig. 2: Induction of gene expression in SURE after addition of subtilin to the culture (P = Phosphorylation)

Multiple expression vectors are available for the SURE system:

  • pNZ8901 (PBS031) - SURE expression vector, PspaSmut, CmR; high promoter activity, but some leakage; suitable for the production of non-toxic proteins. 
  • pNZ8911 (PBS032) - SURE expression vector, PspaS, CmR; no promoter activity without subtilin induction, lower expression than PspaSmut; suitable for the production of potentially toxic proteins. 
  • pNZ8902 (PBS033) - SURE expression vector, PspaSmut, EmR; high promoter activity, but some leakage; suitable for the production of non-toxic proteins. 
  • pNZ8910 (PBS034) - SURE expression vector, PspaS, EmR; some promoter leakage without subtilin induction, caused by the vector; lower expression than pNZ8911.

Several B. subtilis strains specifically developed for the SURE system are available as well. Additionally, E. coli strain MC1061 can be used with the SURE system.

 

Please note these products require a license for non-academic institutes. Please contact us for licensing information.

Features

  • Tightly controlled gene expression in a widely used bacterium 
  • Genome sequence available 
  • Longstanding genetic engineering experience 
  • Complete set of genetic engineering tools available 
  • Potentially useful for cloning of genes with toxic products 
  • Can be used for the identification of essential genes after insertion into the chromosome upstream of the gene in question 
  • Potential for secretion of gene products 
  • Controlled gene expression for metabolic engineering

 

Contents

  • PBS031 - pNZ8901 vector, lyophilized plasmid DNA, 10 µg
  • PBS032 - pNZ8902 vector, lyophilized plasmid DNA, 10 µg
  • PBS033 - pNZ8910 vector, lyophilized plasmid DNA, 10 µg
  • PBS034 - pNZ8911 vector, lyophilized plasmid DNA, 10 µg

 

Literature

 

Publications