| abstract |
Disclosed is useful construct and method, is used for protein expression, uses the primary translation product of processing in the recombinant host cell.Describe the construct that comprises single open reading frame (sORF), be used for protein expression, comprised a plurality of polypeptide expression.Primary translation product (precursor protein or polyprotein) contains polypeptide, and from body processing structure territory, or its variant inserts between a plurality of target protein subunits in the frame as albumen intron or hedgehog family.Elementary product can also contain cleavage sequence, as other proteolysis cracking or proteolytic enzyme recognition site, or contains the signal peptide of signal peptidase recognition sequence, separates at least two in a plurality of protein protomers.The sequence from body processing polypeptides or cracking site that can handle insertion improves the expression efficiency of a plurality of protein protomers separately.The effective expression that carries out protein such as immunoglobulin (Ig) is also disclosed, the independent aspects of secretion and/or the assembling of many subunits.Polyprotein contain heavy chain immunoglobulin and light chain segments or can the segmental situation of antigen recognition in, in one embodiment, selectable stoichiometric ratio is the copy of at least two light chain segments of each heavy chain fragment, and the result is the generation that has formed the correct function antibody that folds and assemble.Described the signal peptide of modifying, comprised from those of light chain immunoglobulin. |