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
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 pPT7 (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 (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 carries two resistances to ampicillin (for E .coli) and tetracycline (for B. megaterium).
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.
order #
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description
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amount
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BMEGT702
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Bacillus megaterium protoplasts, strain MS941, pretransformed with pT7-RNAP
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5 x 500 µL
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BMEGT701
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Bacillus megaterium high yield T7 gene expression kit,
includes pretransformed protoplasts BMEGT702 (5 x 500µl), pPT7 cloning vector and pPT7-GFP control vector (vectors lyophilized, 10 µg each)
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Kit
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BMEGT710
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Bacillus megaterium pPT7 cloning vector, lyophilized
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10 µg
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BMEG50
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Bacillus megaterium protoplasts, strain MS941
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5 x 500 µl
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shipped at RT, protoplasts and kit shipped on dry ice
store lyophilized vectors at 4 °C, reconstituted vectors at -20 °C, protoplasts at -70 °C
Vectors are E. coli / B. megaterium shuttle vectors
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