B. megaterium T7 RNA Polymerase High Yield Gene Expression

The first and only T7 RNAP High Yield Gene Expression System for Bacillus megaterium!

The T7 RNAP High Yield Gene Expression System Features

• Tightly regulated and efficiently inducible xylA operon/T7 RNA polymerase
• Strong T7 RNAP promoter with unique sequence
• Stable, high yield protein production up to 8-fold increased in comparison to common xylose inducible expression
• Easy transformation by use of pretransformed B. megaterium protoplasts
• Control vector with GFP-sequence included in the kit

High Yield Gene Expression with our new T7 RNA Polymerase Expression System!

This system combines the benefits of the tightly regulated and strong T7 RNA polymerase expression system and alkaline protease free Bacillus megaterium. Our system is based on two parallel-replicating plasmids: pT7-RNAP and pP_T7 (Gamer et al. 2009). In addition to the t7 rnap gene under control of the strong xylA promoter pT7-RNAP contains the genes of ampicillin and chloramphenicol for easy selection in E. coli (Amp^R) and B. megaterium (Cm^R).

pP_T7 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 carries two resistances to ampicillin (for E .coli) and tetracycline (for B. megaterium).

Fig. 1 Comparison of GFP amounts produced by variant expression systems. GFP produced by B. megaterium carrying the common xylose-regulated Plasmide pRBBm34 (square), B. megaterium transformed with the T7 expression system plasmids (circle) and B.megaterium with the T7 expression system plasmids treated with rifampicin. (Gamer et al., 2009)

This film shows a growing micro-colony of cells of Bacillus megaterium MS941 expressing GFP observed in a Zeiss Axiovert 200 M microscope. The cells are induced shortly before the film starts. A brightfield image and a fluorescence image are recorded every 5 minutes over a timespan of 5 1/2 hours. For this film the brightfield images are overlayed by the green colored fluorescence images. Recorded by Stefan Leupold, Institut für Mikrobiologie, Technische Universität Braunschweig.

Pretransformed B. megaterium protoplasts!
For your convenience we offer B. megaterium protoplasts pretransformed with pT7-RNAP, so that you just have to insert your gene of interest into pPT7 and transform the pretransformed protoplasts with this plasmid. For control purposes the GFP-expressing vector pPT7-GFP is included in the kit.

Kit Components
Bacillus megaterium high yield T7 gene expression kit, includes pretransformed protoplasts BMEGT702 (5 x 500µl), pP_T7 cloning vector and pP_T7-GFP control vector (vectors lyophilized, 10 µg each)

Download Our T7 RNA Polymerase Expression System Handbook

NOTE: Please note that protoplasts of Bacillus megaterium have a limited shelf life and therefore purchase and use of protoplasts must be thoroughly planned.