I want to generate periplasmic protein of another bacteria in E. coli BL21 strain ? How can i do this ? I verified the cloning with pcr but protein production is not happening. Anyone have a recommendation? Thanks a lot.
Phani Kashyap S
Hi Sena Nur
Have you done insilico characterization of gene of interest.
Confirm the clone with restriction enzyme.
And explain brief about the protocol following for protein expression
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Abul Tarafder
For periplasmic expression you will need a leader sequence to direct it to the periplasm. Have you cloned the protein of interest leader sequence into your construct? Is it one that will be recognised in E.coli? If not you may want to add the pelB leader sequence to the N-terminus of your construct.
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