problems with inclusions bodies

Parmentier Fabrice parmentf at ERE.UMontreal.CA
Mon Feb 21 11:58:46 EST 1994


Hi netters,

Recently I did a construction using the GST gene fusion system with a hydrophobic peptide of 40 amino acids. This fusion protein was well expressed in E.Coli but was unfortunately associated with the pellet fraction and probably localized in inclusion bodies. I have tried to alter the growth conditions of the bacteria but in each case the fusion protein still associated with the pellet fraction. I have tried to solubilize the inclusion bodies of the pellet fraction with: CHAPS 1%, Triton X-100,1%, urea or Gn-HCl and every time the fusion protein is still present in the pellet fraction. I could try a more drastic treatment of this fraction. However, I will need to purify the fusion protein with glutathione Sepharose 4B beads.

Any suggestions or comments are appreciated

Isabelle Lemire
parmentf at ere.umontreal.ca
 





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