Bacillus megaterium hp-Vector (High Performance)

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.

High Performance B. megaterium Vectors

• p3STOP1623hp (BMEG30) - contains two additional stop codons downstream of the multiple cloning site, an optimized -35 region of P_xylA 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.

• pSP_LipA-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. 

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

• p3STOP1623-2RBShp (BMEG35) - carries an additional optimized ribosomal binding site. It is suitable for simultaneous dual expression.

• pC-STREP1623hp (BMEG36) - allows a C-terminal StrepII-tag fusion to the protein of interest.

• pN-STREP-Xa1623hp (BMEG37) - encodes an N-terminal StrepII-tag fusion followed by a Factor Xa recognition site which is cleavable with Factor Xa protease.

• pN-STREP-TEV1623hp (BMEG38) - carries the coding region of an N-terminal StrepII-tag fusion with TEV (tobacco etch virus) recognition site which is cleavable with TEV protease.

• pMGBm19 (BMEG39) - E.coli/Bacillus shuttle vector with xylose-inducible P_xylA promoter designed for co-expression studies. 

• pGFP1624hp (BMEG40C) - positive control vector for GFP expression in B. megaterium.

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