Methods and compositions for expression of polypeptides in a cell

Abstract

Disclosed herein are vector systems for expression of polypeptides in eukaryotic cells; and methods of obtaining high-level expression of polypeptides in a eukaryotic cell. Methods and compositions for obtaining stable, long-term expression of recombinant polypeptides are also provided

What is claimed is: 
     
         1 . A polynucleotide comprising a viral promoter, a transgene, a polyadenylation signal, and a post-transcriptional element (PRE) selected from the group consisting of a woodchuck hepatitis virus PRE (WPRE), a human hepatitis B virus PRE (HPRE), and hybrids thereof, wherein the polynucleotide does not include viral untranslated region (UTR) or viral intron A. 
     
     
         2 . The polynucleotide of  claim 1 , wherein the polyadenylation signal is selected from the group consisting of the simian virus 40 (SV40) polyadenylation signal and the bovine growth hormone (BGH) polyadenylation signal. 
     
     
         3 . The polynucleotide of  claim 1 , further comprising a multiple cloning site. 
     
     
         4 . The polynucleotide of  claim 1 , further comprising a MAR/SAR sequence. 
     
     
         5 . The polynucleotide of  claim 1 , wherein the MAR/SAR sequence is selected from the group consisting of a chicken lysozyme MAR/SAR (CLM) sequence, an interferon alpha-2 MAR/SAR (IAM) sequence, an interferon beta MAR/SAR (IBM) sequence, a X29 MAR/SAR sequence, a S4 MAR/SAR sequence and hybrids thereof. 
     
     
         6 . The polynucleotide of  claim 1 , wherein the transgene is a gene that encodes a protein selected from the group consisting a recombinant protein, a fusion protein, an antibody, a cytokine, a hormone, an enzyme and a clotting factor. 
     
     
         7 . A cell comprising the polynucleotide of  claim 1 , wherein the cell is selected from the group consisting of a prokaryotic cell and a eukaryotic cell.