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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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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 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. BMEG protoplasts ready for transformation can be ordered separately.

Multiple B. megaterium shuttle vectors are available, as well as several control vectors. The shuttle vectors are divided into two series:

  1. Vectors of the 1520 series – have far-off located stop codon (more than 120 bp downstream of start codon) or a stop codon directly downstream of an existing C-terminal tag.
  2. Vectors of the 1622 series – size-reduced vector variants with a closely located stop codon directly downstream of the MCS or a stop codon directly downstream of an existing C-terminal tag.


Vectors of the 1520 series:

  • pWH1520 (BMEG03) - original shuttle vector of the 1520 series. All BMEG vectors, with the exception of pMGBm19, were derived from pWH1520.

  • pMM1522 (BMEG10) – version of pWH1520 with additional restriction site BsrGI upstream of the ATG. The BsrGI site can be used to clone the gene of interest with its native start codon (corresponding to the native N-terminus of the protein of interest). The vector contains a stop codon downstream of the ATG (> 120 bp).

  • pMM1525 (BMEG11) – secretion vector; version of pMM1522 with additional signal sequence for protein secretion (SPlipA) upstream of the MCS (the native lipA encodes for an extracellular esterase).

  • pHIS1522 (BMEG12) – version of pMM1522 with additional sequence for C-terminal 6xHis-tag fusion (including stop codon).

  • pHIS1525 (BMEG13) – secretion vector; version of pMM1525 with additional sequence for C-terminal 6xHis-tag fusion (including stop codon).

  • pSTREP1525 (BMEG14) – secretion vector; version of pMM1525 with additional sequence for N-terminal Strep-tag fusion (cleavable with Factor Xa protease) and stop codon downstream of the MCS; does not contain BsrGI restriction site.

  • pSTREPHIS1525 (BMEG15) – secretion vector; version of pMM1525 with additional sequences for N-terminal Strep-tag (cleavable with Factor Xa protease) and C-terminal His-tag fusion (including stop codon); does not contain a BsrGI restriction site.

  • pC-His1622 (BMEG20) - size-reduced pHIS1522 variant with sequence for C-terminal 6xHis-tag fusion (including stop codon right downstream of the tag).

  • pC-Strep1622 (BMEG21) - size-reduced pHIS1522 variant with sequence for C-terminal StrepII-tag fusion (including stop codon directly downstream of the tag).


Vectors of the 1622 series:


  • pC-His1622 (BMEG20) - size-reduced pHIS1522 variant with sequence for C-terminal 6xHis-tag fusion (including stop codon right downstream of the tag).

  • pC-Strep1622 (BMEG21) - size-reduced pHIS1522 variant with sequence for C-terminal StrepII-tag fusion (including stop codon directly downstream of the tag).

  • pN-His-TEV1622 (BMEG22) - version of pSTOP1622 with an additional sequence for N-terminal 6xHis-tag fusion; cleavable with TEV (tobacco etch virus) protease.

  • pN-Strep-TEV1622 (BMEG23) - version of pSTOP1622 with an additional sequence for N-terminal fusion of StrepII-tag; cleavable with TEV (tobacco etch virus) protease. 

  • pN-StrepXa1622 (BMEG24) – version of pSTOP1622 with additional sequence for N-terminal fusion of StrepII-tag; cleavable with Factor Xa protease.

  • pSTOP1622 (BMEG25) - size-reduced pMM1522 variant with a stop codon directly downstream of the MCS.


Vector

Cat. #

Tag(s)

Secretion

pWH1520

BMEG03


 

pMM1522

BMEG10

 

 

pMM1525

BMEG11

Yes

pHIS1522

BMEG12

6xHis C-terminal tag

pHIS1525

BMEG13

6xHis C-terminal tag

Yes

pSTREP1525

BMEG14

Strep Tag (N-term)

Yes

pSTREPHIS1525

BMEG15

6xStrepII N-terminal tag (cleavable), 6xHis C-terminal tag

Yes

pC-His1622

BMEG20

6xHis C-terminal tag

pC-Strep1622

BMEG21

6xStrepII C-terminal tag

pN-His-TEV1622

BMEG22

6xHis N-terminal tag, cleavable

 

pN-Strep-TEV1622

BMEG23

6xHis StrepII N-terminal tag, cleavable

 

pN-StrepXa1622

BMEG24

6xStrepII N-terminal tag, cleavable

pSTOP1622

BMEG25

 


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

Features

  • Stable, high-yield protein expression 
  • Ideal for both small- and large-scale protein production
  • Tightly regulated and efficiently inducible xylose operon 
  • No alkaline proteases activity even up to 5 hours after induction
  • No endotoxins observed
  • Versatile cloning due to extended polylinker and additional BsrGI site
  • Versatile production (intracellular or extracellular)
  • Versatile purification (native, 6xHis-tag, Strep-tag, Strep/6xHis double-tag)
  • Removable tag versions for TEV or Xa proteases are available

Applications

  • Host for the expression of non-homologous DNA

Vectors

  • BMEG10 - pMM1522 vector, lyophilized DNA, 10 µg
  • BMEG11 - pMM1525 vector, lyophilized DNA, 10 µg
  • BMEG12 - pHIS1522 vector, lyophilized DNA, 10 µg
  • BMEG13 - pHIS1525 vector, lyophilized DNA, 10 µg
  • BMEG14 - pSTREP1525 vector, lyophilized DNA, 10 µg
  • BMEG15 - pSTREPHIS1525 vector, lyophilized DNA, 10 µg
  • BMEG20 - pC-His1622 vector, lyophilized DNA, 10 µg
  • BMEG21 - pC-Strep1622 vector, lyophilized DNA, 10 µg
  • BMEG24 - pN-StrepXa1622 vector, lyophilized DNA, 10 µg
  • BMEG25 - pSTOP1622 vector, lyophilized DNA, 10 µg

Other available BMEG vectors: BMEG03, BMEG22, BMEG23

Literature



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