Innovative Tools for Molecular and Cell Biology

Innovative Tools for Molecular and Cell Biology

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SURE® Expression System for Bacillus subtilis

The SURE® System: Subtilin Regulated gene Expression

Components of the SURE system

Fig. 1 Components of the SURE system

Induction of gene expression in SURE after addition of subtilin to the culture

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

Features

  • Tightly controlled gene expression in a bacterium with long history of biotechnology
  • Genome sequence available
  • Longstanding genetic engineering experience
  • Complete set of genetic engineering tools available
  • Subtilin producing strain cuts costs
  • Potentially useful for cloning of genes with toxic products
  • Potential for secretion of gene products
  • Controlled gene expression for metabolic engineering
  • All vectors are B. subtilis/E. coli shuttle vectors

Description

Controlled gene expression in B. subtilis – a cost-saving alternative to IPTG induced gene expression

  • IPTG (Isopropyl-ß-D-thiogalactopyranoside) is a widespread, established, and non-degradable inducer. However, since IPTG is a considerably expensive compound, an inexpensive method for induction is desirable.

  • Subtilin is 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.
    For the SURE system the 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). Figure 1 shows the basic components of the SURE system.
  • Upon addition of subtilin the system is activated and the protein of interest is produced. It accumulates either intracellularly (blue), or is secreted (green) into the medium (see Figure 2).

Resources

  • The B. subtilis centered wiki SubtiWiki: A community-curated consensual annotation that is continuously updated.
  • SubtiPathways is a model of B. subtilis metabolism and regulation in SBML/SBGN (Systems Biology Markup Language/ Graphical Notation).

ORDER INFORMATION

  • For shipping and storage information please click on Order#.
Order# Description Amount Price Data Sheet
VS-ELS10610-01 NICE E. coli host Strain MC1061 1 ml 305,00 PDF
VS-ELS10710-01 MC1061 Chemically Competent Escherichia coli 5 x 100 µl 438,00 PDF
PBS023 Bacillus subtilis strain NZ8963 1 ml 529,00 PDF
PBS024 Bacillus subtilis strain NZ8900 1 ml 529,00 PDF
PBS025 Bacillus subtilis strain NZ8901 1 ml 350,00 PDF
PBS031 pNZ8901 vector, lyophilized DNA 10 µg 529,00 PDF
PBS032 pNZ8902 vector, lyophilized DNA 10 µg 529,00 PDF
PBS033 pNZ8910 vector, lyophilized DNA 10 µg 529,00 PDF
PBS034 pNZ8911 vector, lyophilized DNA 10 µg 529,00 PDF
  • All prices are in EURO excl. VAT and shipping. For further pricing and order information please ask your local distributor.

Download

The SURE Gene Expression System for Bacillus subtilis - Handbook (PDF)

Vector maps and Sequences

Vector map of pNZ8901 (pic)
Vector map of pNZ8902 (pic)
Vector map of pNZ8910 (pic)
Vector map of pNZ8911 (pic)

Sequence of pNZ8901 (txt)
Sequence of pNZ8902 (txt)
Sequence of pNZ8910 (txt)
Sequence of pNZ8911 (txt)

Literature

  • Bongers, R. S., J. W. Veening, M. Van Wieringen, O. P. Kuipers, and M. Kleerebezem. 2005. Development and characterization of a subtilin-regulated expression system in Bacillus subtilis: strict control of gene expression by addition of subtilin. Applied and Environmental Microbiology 71:8818-8824. Pubmed
  • Kleerebezem, M. 2004. Quorum sensing control of lantibiotic production; nisin and subtilin autoregulate their own biosynthesis. Peptides 25:1405-1414. Pubmed
  • Kleerebezem, M., R. Bongers, G. Rutten, W. M. de Vos, and O. P. Kuipers. 2004. Autoregulation of subtilin biosynthesis in Bacillus subtilis: the role of the spa-box in subtilin-responsive promoters. Peptides 25:1415-1424. Pubmed
  • Mierau, I., and M. Kleerebezem. 2005. 10 years of the nisin-controlled gene expression system (NICE) in Lactococcus lactis. Applied Microbiology and Biotechnology 9:1-13. Pubmed
  • Vavrova, L., Muchova, K., and Barák, I. 2010. Comparison of different Bacillus subtilis expression systems.
    Res Microbiol. 161(9):791-7. Pubmed