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B. megaterium Protein Expression

BMEG VECTORS - Bacillus megaterium shuttle vectors, 10μg

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B. megaterium vector



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PRODUCT DESCRIPTION:
Our Bacillus megaterium system offers an interesting alternative to the standard host E. coli. A variety of E. coli / Bacillus megaterium shuttle vectors with different features (see below) are available. B. megaterium protoplasts ready for transformation can be ordered separately.

B. megaterium has proven to be an excellent host for the expression of non-homologous DNA. It has the advantage over other bacilli strains that none of the alkaline proteases are present. This fact enables cloning and expression of foreign proteins without degradation. In addition, there are no endotoxins found in the cell wall. Protein yields are exceptionally good, even if inexpensive substrates are used. Using the tightly regulated xylose operon the genes are 130- to 350-fold induced without proteolysis. An excellent polylinker downstream of the promoter allows versatile cloning in these vectors.

FEATURES:

  • Stable protein expression with high yield
  • Ideal for both small and large scale protein production
  • Xylose Operon: Tightly regulated and efficiently inducible by xylose (up to 350-fold)
  • Extended polylinker downstream of promoter allows versatile cloning
  • Additional BsrGI site enables cloning without modifying the N-terminus
  • no indication of proteolytic instability even up to 5 hours after induction, since alkaline proteases (such as e.g. in B. subtilis) are not produced
  • Endotoxins are not found in the cell wall
  • Versatile production (intracellular or extracellular)
  • Versatile purification (native, 6xHis-Tag, Strep-Tag, 6xHis/Strep double-tag)
  • Removable tag version (for Xa protease) available for Strep-Tag
  • Suited for industrial large scale protein production as well as laboratory research
  • All B. subtilis vectors are compatible

bmeg

Summary of available B. Megaterium vectors: (NOTE: All 1622 series vectors lack 855 superfluous base pairs compared to the 1522 series)

VectorCat. #TagSecretion
pMM1522BMEG10NONO (intracellular)
pMM1525BMEG11NOYES (extracellular)
pHIS1522BMEG126 X HIS (C-term)NO
pHIS1525BMEG136 X HIS (C-term)YES
pSTREP1525BMEG14Strep Tag (N-term)YES
pSTREPHIS1525BMEG15Strep (N-term)/His (C-term) Double TagYES
pC-His1622BMEG206 X HIS (C-term)NO
pC-Strep1622BMEG21Strep Tag (C-term)NO
pN-StrepXa1622BMEG24Strep Tag (N-term) (Xa site)NO

Proteins successfully over-produced with this system are:

  • Catabolite control protein (ccpA)
  • Xylose repressor (XylR)
  • Trehalose repressor (TreR)
  • Heat shock protein (HPr) from PTS (phosphotransferase sugar transport system)
  • Mutarotase (Mro)
  • Glucose dehydrogenase (Gdh)
  • Beta-galactosidase
  • Human single-chain urokinase-like plasminogen activator (rscuPA)
  • Clostridium difficile toxin A
  • Cobaltochelatase (CbiX)
  • Dextransucrase
  • Endolevanase (LevB)
  • Levansucrase
  • Neopullanase
  • Translocation ATPase of the preprotein translocase (SecA)

PACKAGING: 10ug of vector, lyophilized

Click Here For User Manual
B. megaterium protoplasts ready for transformation can be ordered separately Click Here
For hp (high performance) Bacillus megaterium cloning vectors Click Here


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