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B. subtilis Protein Expression

BSUB VECTORS - Bacillus subtilis vectors, 10μg


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Bacillus subtilis vectors

Gram-positive bacteria are well known for their contributions to agricultural, medical and food biotechnology and for the production of recombinant proteins. Among them, Bacillus subtilis has been developed as an attractive host because of several reasons:

  • It is non-pathogenic and is considered as a GRAS organism (generally regarded as safe);
  • it has no significant bias in codon usage;
  • it is capable of secreting functional extracellular proteins directly into the culture medium (at present, about 60% of the commercially available enzymes are produced by Bacillus species);
  • a large body of information concerning transcription, translation, protein folding and secretion mechanisms, genetic manipulation and large-scale fermentation has been acquired.

But there are also two obstacles reducing the use of B. subtilis: (i) production of a number of extracellular proteases which recognize and degrade heterologous proteins, and (ii) stable vector plasmids. The first obstacle has been largely solved by the construction of protease-deficient strains. And the second has been completely overcome by introducing plasmids using the theta-mode of replication such as those derived from the natural plasmids pAMβ1 and pBS72 (Janničre et al., 1990; Titok et al., 2003).

The new vector pHT01 allows high-level expression of recombinant proteins within the cytoplasm, whereas pHT43 secretes the recombinant proteins into the medium.

Derivatives of pHT01 are available either with a 8xHis tag (pHT08), a Strep tag (pHT09) or a c-Myc tag (pHT10).

pHT01 pHT43

The vectors are based on the strong σA-dependent promoter preceding the groE operon of B. subtilis which has been converted into an efficiently controllable (IPTG-inducible) promoter by addition of the lac operator.

Summary of available B. Subtilis vectors:

VectorCat. #TagSecretion
pHT01PBS001NONO (intracellular)
pHT43PBS002NOYES (extracellular)
pHT09PBS004Strep TagNO
pHT10PBS005c-Myc TagNO

Please note, a license is required for PBS001, PBS002, PBS003, PBS004, and PBS005 for any non-academic customer.  Please contact us at info@bocascientific.com for more information.

Bacillus subtilis User Manual