Multiple gene expression including sorf contructs and methods with polyproteins, pro-proteins, and proteolysis

摘要

Disclosed is an expression vector for generating one or more recombinant protein products comprising a promoter regulatory element operably linked to a single open reading frame (sORF) insert; said sORF insert comprising: (a) a first nucleic acid sequence encoding a first polypeptide, (b) a first intervening nucleic acid sequence encoding a first protein cleavage site, and (c) a second nucleic acid sequence encoding a second polypeptide; wherein said intervening nucleic acid sequence encoding said first protein cleavage site is operably positioned between said first nucleic acid sequence and said second nucleic acid sequence; and wherein said expression vector is capable of expressing a sORF polypeptide cleavable at said first protein cleavage site; wherein said first protein cleavage site comprises a self-processing cleavage site comprising a hedgehog segment or modified hedgehog segment, wherein the hedgehog segment or modified hedgehog segment permits cleavage of said sORF polypeptide but not ligation of said first polypeptide and said second polypeptide.