I would like to express some short, ~20 residue, ~1.2 kDa oligopeptides in E. coli where they will have an in vivo function. I do *not* need to purify them (which seems to be the concern of most other small protein-related questions on Researchgate).
The sequence is fairly hydrophobic with a net charge of only -1 and Innovagen's peptide solubility calculator thinks they will have "poor water solubility." That said, the peptides should get covalently conjugated to a soluble protein after expression. My questions:
1) Will something this small even be expressed in E. coli?
2) If yes, will it go straight to inclusion bodies or might the peptides hang around long enough to form their covalent adducts? The covalent linkage is a fairly quick reaction (SpyTag/SpyCatcher technology...goes to near completion by ~15' in vitro)
3) I understand SUMOylating the peptides could help their soluble expression and I can do in vivo cleavage via Ulp co-expression but per question 2, will the cleaved peptides then just aggregate?
Thanks for any advice!
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