Patent ID: 8426167

Claim:
A method of producing a fusion polypeptide, comprising: culturing a host cell comprising an expression vector comprising a recombinant DNA molecule encoding the fusion polypeptide, the recombinant DNA molecule comprising a first nucleotide sequence coding for a target polypeptide and upstream to the first nucleotide sequence a second nucleotide sequence coding for an FKBP chaperone selected from the group consisting of FkpA and SlyD, wherein the FkpA chaperone is an active FkpA chaperone consisting of amino acids 26-270 of an inactive precursor wild-type FkpA chaperone, wherein the SlyD chaperone is a C-terminally truncated wild-type SlyD chaperone, wherein the FKBP chaperone provides for a functional chaperone activity, and wherein the FKBP chaperone allows for a cytosolic expression and in vitro refolding of the fusion polypeptide; expressing the fusion polypeptide in the cytosol as inclusion bodies comprising the fusion polypeptide; solubilizing the inclusion bodies comprising the fusion polypeptide; purifying the fusion polypeptide; and refolding the fusion polypeptide.