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B. megaterium Protein Expression

BMEG VECTORS-hp - Bacillus megaterium hp-shuttle vectors (high performance), 10μg

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B. megaterium vector-hp



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The Bacillus 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 host for the expression of non-homologous DNA. Unlike other bacilli strains, none of the alkaline proteases are present. This enables cloning and expression of foreign proteins without degradation. In addition, there are no endotoxins found in the cell wall. B. megaterium can stably maintain several extra-chromosomal DNA elements in parallel. Protein yields are exceptionally high, even if inexpensive substrates are used.

The high-performance (hp) BMEG vectors offer yields up to 10-fold higher than the basic plasmids. The hp vectors include vectors carrying two different signal peptide sequences, a 6xHis-tag, a Strep-tag, or two ribosome binding sites for simultaneous dual expression. Vectors encoding C- or N-terminal His-tags for easy purification are also offered. The protein secretion with LipA or YocH signal peptides is up to 9-fold increased. All hp BMEG vectors have established multiple cloning sites for versatile cloning. Induction of protein expression is achieved by the tightly regulated and efficiently inducible xylose operon.


Multiple hp BMEG vectors are offered:

  • p3STOP1623hp (BMEG30) – contains two additional stop codons downstream of the multiple cloning site, an optimized -35 region of PxylA and an optimized ribosome binding site. All following plasmids are based on this plasmid.

  • pC-His1623hp (BMEG31) - allows a C-terminal 6xHis-tag (3’ of MCS) fusion including a stop codon downstream of the tag.

  • pN-His-TEV1623hp (BMEG32) - encodes an N-terminal 6xHis-tag fusion including a TEV (tobacco etch virus) recognition site that is cleavable with TEV protease treatment.

  • pSPLipA-hp (BMEG33) - contains a His6-tag for N-terminal fusion of recombinant gene including the coding sequence of the signal peptide of B. megaterium protein LipA for secretion.

  • pSPYocH-hp (BMEG34) - encodes the signal peptide of B. megaterium protein YocH that enables secretion of proteins fused to an N-terminal His6-tag.

Vector

Cat. #

Tag

Secretion

p3STOP1623hp

BMEG30

NO

NO

pC-His1623hp

BMEG31

6 x HIS (C-term)

NO

pN-His-TEV1623hp

BMEG32

6 X HIS (N-term) (TEV-site)

NO

pSPLipA-hp

BMEG33

HIS

YES

pSPYocH-hp

BMEG34

HIS

YES



Please note these products require a license for non-academic institutions. Please contact us for licensing information.


Features

  • High performance vectors with optimized sequences
  • Ideal for both small- and large-scale protein production
  • Tightly regulated and efficiently inducible xylose operon (up to 350-fold)
  • No alkaline proteases activity even up to 5 hours after induction
  • No endotoxins observed
  • Multiple cloning sites for versatile cloning
  • Encoding C- or N-terminal His-tag for versatile purification (native, 6xHis-tag)
  • Vectors with two ribosome binding sites for simultaneous dual expression are available

Applications

  • Host for the expression of non-homologous DNA

Vectors

  • BMEG30 - p3STOP1623hp vector, lyophilized DNA, 10 µg
  • BMEG31 - pC-His1623hp vector, lyophilized DNA, 10 µg
  • BMEG32 - pN-His-TEV1623hp vector, lyophilized DNA, 10 µg
  • BMEG33 - pSPLipA-hp vector, lyophilized DNA, 10 µg
  • BMEG34 - pSPYocH-hp vector, lyophilized DNA, 10 µg

Literature