Bacillus megaterium High Yield T7 Gene Expression Kit

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 pP_T7. pT7-RNAP contains the genes for ampicillin and chloramphenicol resistance for easy selection in E. coli (Amp^R) and B. megaterium (Cm^R), in addition to the t7 rnap gene under control of the strong xylA promoter. 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, pP_T7 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. 

This kit includes strain MS941, pP_T7 cloning vector, and pP_T7-GFP control vector. A secretion vector, pP_T7-SP_LipA, is separately available. For added convenience, strain MS941 comes pretransformed with pT7-RNAP. This allows the user to immediately insert the gene of interest into pP_T7 and transform the pretransformed protoplasts with this plasmid.

Please note this product requires a license for commercial companies. Please contact us for licensing information.