Vector Bacillus subtilis

Wu, S.C. and Wong, S.L. (1999) Development of improved pUB 110-based vectors for expression and secretion studies in Bacillus subtilis. Journal of Biotechnology, 72 (1-2), 185-195. 

Ferreira, L.C., Ferreira, R.C. and Schumann, W. (2005) Bacillus subtilis as a tool for vaccine development from antigen factories to delivery vectors. Anais da Academia Brasileira de Ciencias, 77 (1), 113-124. 

We routinely use a mix of five mRNAs that are derived from the lys (ATCC no. 87482), trp (ATCC no. 87485), dap (ATCC no. 87486), thr (ATCC no. 87484), andphe (ATCC no. 87483) clones from the bacterium Bacillus subtilis cloned into a vector that contains a stretch of As. These RNAs are generated by in vitro transcription using a T3 in vitro transcription kit (e.g., MEGAscript from Ambion) of the linearized DNA template with the appropriate restriction enzyme. 

Schweizer HP. 1992. Allelic exchange in Pseudomonas aeruginosa using novel ColEl-type vectors and a family of cassettes containing a portable oriT and the counter-selectable Bacillus subtilis sacB marker. Mol Microbiol 6 1195-204. 

The host bacteria used for production of recombinant proteins are usually E. coli, or Bacillus subtilis they may express proteins at 1 % to over 50% of the cellular protein, depending on such variables as the source, promoter structure, and vector type. Generally the proteins are expressed intracellularly, but leader sequences for excretion may be included. In the latter case, the protein is generally excreted into the periplasmic space, which limits the amount that can be produced. Excretion from grampositive species such as B. subtilis sends the product into the culture medium, with little feedback limitation on total expression level. 

V Vagner, E Dervyn, SD Ehrlich. A vector for systematic gene inactivation in Bacillus subtilis. Microbiology 144 3097-3104, 1998. 

Bacillus subtilis Many proteins can be exported into growth medium Easy to grow in large-scale volumes Regulation of gene expression not very well known Lack of high level expression vectors Unable to carry out most post-translational modifications, e.g. glycosylation... 

Clostridium acetobutylicum Acetone and butanol production Overexpression of acetoacetate decarboxylase (adc) and phosphotransbutyrylase (ptb) by introducing a Bacillus subtilis/C acetobutylicum shutde vector into C. acetobutylicum by an improved electrotransformation protocol, which resulted in acetone and butanol formation 88... 

New shuttle vectors for ectopic insertion of genes into Bacillus subtilis. Plasmid, 51 (3), 238-245. 

Chary, V.K., Amaya, E.I., and Piggot, P.J. (1997) Neomycin- and spectinomycin-resistance replacement vectors for Bacillus subtilis. FEMS Microbiol Lett, 153 (1), 135-139. 

E, and Jaeger, K.E. (2012) Novel broad host range shuttle vectors for expression in Escherichia coli. Bacillus subtilis and Pseudomonas putida. J. Biotechnol, 161 (2), 71-79,... 

Appendix E-I-A. Bacillus subtilis Host-Vector 1 Systems Appendix E-I-B. Bacillus subtilis Host-Vector 2 Systems Appendix E-II. Saccharomyces cerevisiae... 

A B1 host-vector system consists of a host with low viability in natural conditions. The vector must be dependent on the host and incapable of transfer to other cells. Examples of B1 host-vector systems are EKl (based on the bacterium Escherichia coli K12 and a plasmid which cannot conjugate or transfer to other bacteria), SCI (a laboratory-maintained strain of the yeast Saccharomyces cerevisiae as host and a plasmid or minichromosome as vector) and BSl (the bacterium Bacillus subtilis Marburg 168 as host and a plasmid). B2 host-vector systems consist of a host with especially low viability in natural conditions and a vector which depends on the host completely. The most common example is EK2 (a defective strain of E. coli K12 and the well-characterised plasmid pBR322). 

In the future the use of vectors to clone foreign DNA is certain to be extended to other host/vector partners. The value of less pathogenic bacterial hosts such as Bacillus subtilis has recently been reviewed. Plant viruses as vectors for higher plants hold much promise for the insertion of characters such as disease resistance and nitrogen fixation into plants of agricultural importance. ... 

M. and Tamura, G., 1984, Secretion vector of Bacillus subtilis constructed from the Bacillus subtilis a-amylase promoter and signal peptide coding region, "Genetics and Biotechnology of Bacilli", A.T. Ganasan and J.A. Hoch, eds.. Academic Press, London and Orlando, pp. 181. 

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