I'm cloning a Gram negative bacterial secreted toxin in pET15b and pET28a vectors. I have confusion that which strain of E. coli would be better for induction of protein expression, BL21 or BL21(DE3).
use BL21(DE3),
Clone in DH10B or DH5alpha strains first than isolate plasmid and transform in BL21(DE3) for expression analysis. If the toxin is toxic to E. coli you wont be able to express or even get the BL21(DE3) transformants.
BL21 does not have T7 RNA pol encoding region i.e. DE3. So, it cannot be used for expression of genes cloned downstream to T7 promoter like in pET15b and pET28a.
use BL21(DE3),
Clone in DH10B or DH5alpha strains first than isolate plasmid and transform in BL21(DE3) for expression analysis. If the toxin is toxic to E. coli you wont be able to express or even get the BL21(DE3) transformants.
BL21 does not have T7 RNA pol encoding region i.e. DE3. So, it cannot be used for expression of genes cloned downstream to T7 promoter like in pET15b and pET28a.
Hi
May be, you can try with Lemmo21 , this is modified version of BL21(DE3) and this is specific for transmembrane protein and also for toxic protein.
Good Luck !!
@ Mangal Singh, Michael J. Benedik and @ Shambhu Yadav; Thanks for valuable suggestions, I will try the same.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Molecular Methods
- Cloning