Disclosed are useful constructs and methods for the expression of proteins using primary translation products that are processed within a recombinant host
cell. Constructs comprising a single
open reading frame (sORF) are described for
protein expression including expression of multiple polypeptides. A primary translation product (a pro-
protein or a polyprotein) contains polypeptides such as inteins or
hedgehog family auto-
processing domains, or variants thereof, inserted in frame between multiple
protein subunits of interest. The primary product can also contain cleavage sequences such as other proteolytic cleavage or
protease recognition sites, or
signal peptides which contain recognition sequences for
signal peptidases, separating at least two of the multiple protein subunits. The sequences of the inserted auto-
processing polypeptides or cleavage sites can be manipulated to enhance the efficiency of expression of the separate multiple protein subunits. Also disclosed are independent aspects of conducting efficient expression,
secretion, and / or multimeric
assembly of proteins such as immunoglobulins. Where the polyprotein contains immunoglobulin heavy and light chain segments or fragments capable of
antigen recognition, in an embodiment a selectable stoichiometric ratio is at least two copies of a light
chain segment per
heavy chain segment, with the result that the production of properly folded and assembled functional
antibody is made. Modified
signal peptides, including such from immunoglobulin light chains, are described.