Compositions and methods for enhanced expression of recombinant polypeptides from a single vector using a peptide cleavage site
Vector constructs for expression of two or more functional proteins or polypeptides under operative control of a single promoter and methods of making and using the same are described. The vectors comprise a self-processing cleavage site between each respective protein or polypeptide coding sequence. The vector constructs include the coding sequence for a self-processing cleavage site and may further include an additional proteolytic cleavage sequence which provides a means to remove the self processing peptide sequence from expressed protein(s) or polypeptide(s). The vector constructs find utility in methods for enhanced production of biologically active proteins and polypeptides in vitro and in vivo.
1 - 27 . (canceled)
28 . An oligonucleotide construct consisting essentially of a coding sequence for a furin cleavage site and a coding sequence for a 2A self-processing cleavage site, wherein the coding sequence for the furin cleavage site is located immediately 5′ to the coding sequence for the 2A self-processing cleavage site.
29 . The oligonucleotide construct of claim 28 , wherein the coding sequence for the 2A self-processing cleavage site encodes an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8 and SEQ ID NO: 9.
30 . The oligonucleotide construct of claim 28 , wherein the coding sequence for the 2A self processing cleavage site is SEQ ID NO:14.
31 . The oligonucleotide construct of claim 28 , wherein the coding sequence for the furin cleavage site encodes an amino acid sequence of SEQ ID NO: 10.
32 . The oligonucleotide construct of claim 28 , wherein the coding sequence for the furin cleavage site is SEQ ID NO: 15.
33 . A vector comprising the oligonucleotide construct of claim 28 .