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B. megaterium T7 RNA Polymerase High Yield Gene Expression

BMEGT710 - Bacillus megaterium pPT7 cloning vector, lyophilized, 10ug

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The Bacillus megaterium pPT7 cloning vector, strain MS941, is part of the T7 RNA Polymerase high yield gene expression system for Bacillus 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. 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 Bacillus megaterium high yield T7 gene expression system combines the benefits of the tightly regulated and strong T7 RNA polymerase expression system with the alkaline protease-free Bacillus megaterium. In addition to the t7 rnap gene under control of the strong xylA promoter, pT7-RNAP contains the genes for ampicillin and chloramphenicol resistance for easy selection in E. coli (AmpR) and B. megaterium (CmR). 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, the plasmid comprises two resistances against ampicillin (in E. coli) and tetracycline (in B. megaterium).

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

-Easy transformation by use of pretransformed B. megaterium protoplasts

-Ideal for both small- and large-scale protein production

-Extracellular protein production possible with pPT7-SPlipA

Click Here for User Manual