Innovative Tools for Molecular and Cell Biology

Innovative Tools for Molecular and Cell Biology

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hp-Vector Expression Systems for Bacillus megaterium

High Performance Expression System for B. megaterium

 Bacillus megaterium and Escherichia coli

Electron microscope image of Bacillus megaterium and Escherichia coli, Dr. Manfred Rohde, Helmholtz-Center for Infection Research (HZI), Braunschweig, Germany

Features

  • High performance vectors with optimized sequences
  • Protein yield up to 10 times enhanced compared to protein expression with basic plasmid
  • Plasmids with established MCS
  • Encoding C- or N-terminal His-tag for versatile purification (native, 6xHis-tag)
  • Secretion with LipA or YocH signal peptide up to 9-fold increased
  • Vector with two ribosome binding sites (2RBS) for simultaneous dual expression available
 Relative fluorescence of an optimized hp-Vector (red; square) and a basic plasmid (blue; circle) over time [h].

Relative fluorescence of an optimized hp-Vector (red; square) and a basic plasmid (blue; circle) over time [h].

 Soluble protein fractions 7.5 h after induction of heterologous gene expression.

Soluble protein fractions 7.5 h after induction of heterologous gene expression.

Optimized high performance (hp) vectors for Bacillus megaterium

  • MoBiTec provides a wide range of useful vectors for the B. megaterium system that are adaptable to most applications and protein purification methods. These include vectors carrying two different signal peptide sequences, a 6xHis-tag, a Strep-tag, or two ribosome binding sites (2RBS) for simultaneous dual expression.
  • With our new hp-vectors yields up to 10-fold higher compared to the basic plasmids are possible. All plasmids have established multiple cloning sites (MCS) for versatile cloning. Furthermore, we offer vectors encoding C- or N-terminal His-tags for easy purification. The protein secretion with LipA or YocH signal peptides is up to 9-fold increased. Induction of protein expression of all vectors is achieved by the tightly regulated and efficiently inducible xylose operon.

Versatile system with a wide range of vectors

  • The B. megaterium expression system provides a versatile and easy-to-handle tool for stable and high-yield protein production, both small- and large-scale. B. megaterium has proven to be an excellent alternative host to E. coli for heterologous gene expression. Unlike other bacilli strains, proteolytic degradation by external alkaline proteases is avoided. In addition, there are no endotoxins found in the cell wall.

Order Information

  • For shipping and storage information please click on Order#.
Order# Description Amount Price Data Sheet
BMEG02 Bacillus megaterium protoplasts, strain WH320 5x500 µl 417,00 PDF
BMEG04 Bacillus megaterium protoplasts, strain YYBm1 5x500 µl 480,00 PDF
BMEG50 Bacillus megaterium protoplasts, strain MS941 5x500 µl 464,00 PDF
BMEG30 Bacillus megaterium vector p3STOP1623hp 10 µg 1065,00 PDF
BMEG31 Bacillus megaterium vector pC-HIS1623hp 10 µg 1123,00 PDF
BMEG32 Bacillus megaterium vector pN-HIS-TEV1623hp 10 µg 1123,00 PDF
BMEG33 Bacillus megaterium vector pSP-LipA-hp 10 µg 1169,00 PDF
BMEG34 Bacillus megaterium vector pSP-YocH-hp 10 µg 1169,00 PDF
BMEG35 Bacillus megaterium vector p3STOP1623-2RBShp 10 µg 1100,00 PDF
BMEG36 Bacillus megaterium vector pC-STREP1623hp 10 µg 1141,00 PDF
BMEG37 Bacillus megaterium vector pN-STREP-Xa1623hp 10 µg 1152,00 PDF
BMEG38 Bacillus megaterium vector pN-STREP-TEV1623hp 10 µg 1152,00 PDF
BMEG39 Bacillus megaterium vector, pMGBm19, lyophilized DNA 10 µg 1065,00 PDF
BMEG40C pGFP1624hp, high performance GFP expression vector, positive control 10 µg 532,00 PDF
PEC04 E. coli PxylA repressing vector, pMMEc4, lyophilized DNA 10 µg 215,00 PDF
  • All prices are in EURO excl. VAT and shipping. For further pricing and order information please ask your local distributor.

  • All vectors are E. coli / B. megaterium shuttle vectors.
  • Please note that protoplasts of Bacillus megaterium have a limited shelf life of a few months only, depending on the date of production.
    Therefore, purchase and use of protoplasts must be thoroughly planned.

Downloads

  • hp-Vector Expression Systems for Bacillus megaterium Handbook (PDF)
  • Bacillus megaterium Protein Production System Handbook (PDF)
  • Commentary – A short story about a big magic bug (Link)

Vector maps and Sequences

Vector map of p3STOP1623hp (pic)
Vector map of pC-His1623hp (pic)
Vector map of pN-His-TEV1623hp (pic)
Vector map of pSPLipA-hp (pic)
Vector map of pSPYocH-hp (pic)
Vector map of p3STOP1623-2RBShp (pic)
Vector map of pC-STREP1623hp (pic)
Vector map of pN-STREP-Xa1623hp (pic)
Vector map of pN-STREP-TEV1623hp (pic)
Vector map of pMGBm19 (pic)

Sequence of p3STOP1623hp (txt)
Sequence of pC-His1623hp (txt)
Sequence of pN-His-TEV1623hp (txt)
Sequence of pSPLipA-hp (txt)
Sequence of pSPYocH-hp (txt)
Sequence of p3STOP1623-2RBShp (txt)
Sequence of pC-STREP1623hp (txt)
Sequence of pN-STREP-Xa1623hp (txt)
Sequence of pN-STREP-TEV1623hp (txt)
Sequence of pMGBm19 (txt)

Literature

  • Stammen S, Müller BK, Korneli C, Biedendieck R, Gamer M, Franco-Lara E, Jahn D.: High-yield intra- and extracellular protein production using Bacillus megaterium. Appl. Microbiol. Biotechnol. (2010), 76(12):4037-46. Pubmed
  • Rygus T, Hillen W.: Inducible high-level expression of heterologous genes in Bacillus megaterium using the regulatory elements of the xylose-utilization operon, Appl. Microbiol. Biotechnol. (1991), 35, 594-599. Pubmed
  • Rygus T, Scheler A, Allmansberger R,Hillen W.: Molecular cloning, structure, promoters and regulatory elements for transcription of the Bacillus megaterium encoded regulon for xylose utilization. Arch Microbiol, Jan 1991; 155(6): 535-42. Pubmed
  • Hueck CJ, Kraus A, Schmiedel D, Hillen W.: Cloning, expression and functional analyses of the catabolite control protein CcpA from Bacillus megaterium. Mol Microbiol, Jun 1995; 16(5): 855-64. Pubmed
  • Vary PS.: Prime time for Bacillus megaterium Microbiology, May 1994; 140: 1001. Pubmed
  • Saxena A, Zhang RW, Bollag JM.: Microorganisms capable of metabolizing the herbicide metolachlor. Appl. Environ. Microbiol. (1987) 53, 390-396. Pubmed
  • Selvanayagam, M. and Vijaya, J., Degradation of persistent insecticides by aquatic bacteria as sole source of carbon. J. Environ. Biol. (1989) 10, 399-407.
  • Meinhardt F, Stahl U, Ebeling W. Highly efficient expression of homologous and heterologous genes in Bacillus megaterium. Appl. Microbiol. Biotechnol. (1989) 30, 343-350.PUYET A. (1) ; SANDOVAL H. ; LOPEZ P. ; AGUILAR A. ; MARTIN J. F. ; ESPINOSA M.
  • Puyet A, Sandoval H, Lopez P, Aguilar A, Martin JF, Espinosa M.: A simple medium for rapid regeneration of Bacillus subtilis protoplasts transformed with plasmid DNA. FEMS Microbiol. Lett. (1987) 40, 1-5.
  • Laemmli, U. K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (1970) 227, 680-685 Pubmed
  • Antelmann, H., Tjalsma, H., Voigt, B., Ohlmeier, S., Bron, S., van Dijl, J.M. and Hecker, M. : A proteomic view on genome-based signal peptide predictions. Genome Res. 2001 Sep;11(9):1484-502. Pubmed
  • Malten, M., Biedendieck, R., Gamer, M., Drews, A.-C., Stammen, S., Buchholz, K., Dijkhuizen, L. and Jahn, D.: A Bacillus megaterium plasmid system for the production, export and one-step purification of affinity tagged heterologous levansucrase from the growth medium. Appl Environ Microbiol. 2006 Feb;72(2):1677-9. Pubmed
  • Wang W, Hollmann R, Deckwer WD.: Comparative proteomic analysis of high cell density cultivations with two recombinant Bacillus megaterium strains for the production of a heterologous dextransucrase. Proteome Sci. 2006 Oct 5;4:19. Pubmed

Potential industrial and diagnostical applications

  • Vary, P. S. (1992) Development of genetic engineering in Bacillus megaterium : an example of the versatility and potential of industrially important bacilli Biotechnology, Jan 1992; 22: 251-310. Pubmed
  • Ginsburgh, C., Spaulding, D., Robey, G., Shivakumar, A.M.O., Vanags, R., Katz, L., Fox, J.L. (1989): Sporulation promoter spoVG controlled expression of PP42 gene of HIV-1 in Bacillus megaterium. Abstract from an International Conference on AIDS, Montreal.