hi there, want to ask you what kind of vector that can be used to transform the clone into BL21DE3 for protein expression? Currently im using pET SYSTEM and seem its not working. So i decide to used another system,so what the choice i have?TQ for your respond
Hi there,
The aim of BL21(DE3) strain is to induce with IPTG the expression of T7 polymerase which as a result will highly promotes the transcription of genes of interest under T7 promoter control. So any T7 promoter sequence containing plasmid is appropriate. Furthermore as IPTG is the inducer in this strain any IPTG inducible promoter may also be considered but it would be a bit of a waste as T7 RNA polymerase will be systematically coexpress with the protein of interest...
BL21 DE3 is an expression strain where the over expression of your desired construct is under the control of T7 promoter. So not all vectors will be useful and compatible with BL21 DE3.
The pET family is a very large family of vectors which are popular when working with BL21 DE3. So you could see if shifting vectors within the pET family helps. If not, you might look into pBAT4/ pBAT5/ pCal-n. These vectors incorporate T7 promoters so should be good to work with in BL21 DE3 strain. Please go through the following list from EMBL for a more detailed list.
https://www.embl.de/pepcore/pepcore_services/strains_vectors/vectors/bacterial_expression_vectors/popup_bacterial_expression_vectors/
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