Dear David Vizarraga

i do not know the pOPIN vectors?

Which kind of promoter is present for the protein expression in E.coli?

you can certanilly add pelB or OmpA signal peptide at the N-terminus of the chain. I

Several years go i used periplasmic expression in E.coli to produce a panel of Scfv carryng those signal peptides at N-term and his tag at the C-tem.

This sistem work, the scfv produced were able to bind the target antigens but the expression yields were quite low (

Manuele Martinelli Hello! Thanks for your answer!

I am trying with T7promoter+PelB sequence to express the protein. We compared too with a cytosolic expression.

David

Dear David

if you try also the citosolic expression, use the Origami or the Shuffle E.coli straiins which has less reducing cytoplasm and allow some possibilities to express the with foScfvrmed S-S bonds. In my experience the periplasmic expression work better but it may change on the basis of your scfv sequence and expression/lysis conditions.

best regards

Manuele