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B. megaterium T7 RNA Polymerase High Yield Gene Expression

BMEGT702 - Bacillus megaterium protoplast strain MS941, pretransformed with pT7-RNAP, 5 X 500μl

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The pretransformed Bacillus megaterium protoplast, strain MS941, is part of the T7 RNA Polymerase high yield gene expression system for Bacillus megaterium. This B. megaterium protoplast comes pretransformed with pT7-RNAP so that the gene of interest can be inserted into pPT7 immediately. Please note: this protoplast has a limited shelf life of a few months from the date of production.

About the T7 RNA polymerase expression system

The Bacillus megaterium high yield T7 gene expression system combines the features of the B. megaterium system in E. coli with the T7 RNA polymerase expression system regulation by the xylose operon. This dual plasmid system is based on two parallel-replicating plasmids: pT7-RNAP and pPT7pT7-RNAP contains the genes for ampicillin and chloramphenicol resistance for easy selection in E. coli (AmpR) and B. megaterium (CmR), in addition to the t7 rnap gene under control of the strong xylA promoter. pPT7 is responsible for the T7 RNAP-dependent expression of the target gene. Downstream of the T7 promoter, it comprises a multiple cloning site with ten unique restriction enzyme cleaving sites. Additionally, pPT7 comprises two resistances against ampicillin (in E. coli) and tetracycline (in B. megaterium).

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 and no endotoxins are 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. 

Please note this product requires a license for non-academic institutions. Please contact us for licensing information.

Features

  • Strong T7 RNAP promoter with unique sequence
  • Tightly regulated and efficiently inducible xylA operon/T7 RNA polymerase
  • Suitable for cloning of toxic proteins in E. coli
  • Stable, high yield protein production up to 8-fold increase compared to common xylose inducible expression
  • Ideal for both small- and large-scale protein production

Contents

  • Pretransformed B. megaterium protoplast, strain MS941, 500µl x 5

Literature